Epigenetics Compound Library

Epigenetic chemical compound library screen for the effect of 24 compounds on adipocytic differentiation of human skeletal (mesenchymal) stem cells (hMSCs). Representative Oil Red O staining of lipid-filled mature adipocytes on day 7 after treatment with the indicated compounds (500 nM). Images were taken at 320 magnification using a Zeiss inverted microscope. Abbreviation: DMSO, dimethyl sulfoxide.

Stem Cells Transl Med, 2016, 5(8):1036-47.. purchased from Selleck

Compound screening and confirmation. (A) High-throughput screening of CHIKV 26S mediated insect cell fusion inhibitors obtained from a library of 788 FDA-approved drugs. (B) Microneutralization assay. The square frame represents the protective effects of niclosamide and nitazoxanide against CHIKV-induced CPE. (VC: as CHIKV infection control, CC: as negative control.)

Antiviral Res, 2016, 135:81-90.. purchased from Selleck

(c) Dot plot summarizes the results of Phase I EpI screen. y-Axis indicates cell viability and x-axis represents induction of Ecad promoter activity. Five drugs, Vorinostat, Bortezomib, Etravirine, Niclosamide and Crystal violet, were identified with more than twofold Ecad promoter activity. (d) Dot plot summarizes the results of Phase 2 screen on HDAC inhibitors.

Cell Death Discov, 2016, 2:16041.. purchased from Selleck

Summary of the CCN1-GFs screening, calculated as the relationship between GFP fluorescence intensity using a fluorescence meter.

Cytotechnology, 2016, 68(4):1633-40.. purchased from Selleck

Hit compounds display varying effects on p‐EGFR and EGFR levels. Of 21 EGFR/ERBB hit compounds that selectively targeted chordoma cells, the impact of 13, comprising a selection of hit compounds across the libraries and chemical structures tested, was studied by western blot on three chordoma cell lines (U-CH1, U-CH2, MUG-Chor1). Cells were serum‐starved overnight before being treated with EGFR inhibitors (250 nm) for 4 h and then being exposed to EGF (50 ng/ml) for 15 min.

J Pathol, 2016, 239(3):320-34.. purchased from Selleck

Results of clinical collection compound primary screen. Panel A. Scatterplot representing the results for the screen at 5 μM. Green circles represent the hits, blue circles are all the other compounds. Panel B. Scatterplot representing the results for the screen at 15 μM. Green circles represent the hits from the 5 μM screen, blue circles are all the other compounds. Panel C. Scatterplot comparing T. cruzi percent inhibition between 5 μM and 15 μM screens. Panel D. Replicates plot for a subset of compounds screened twice at 5 μM (n = 579, R2 = 0.86).

PLoS Negl Trop Dis, 2016, 10(4):e0004584.. purchased from Selleck

Graphical representation of hit compounds that block cardiomyocyte hypertrophy. (A) Compounds from the Spectrum Library are depicted based on percent inhibition of ANF expression (X-axis) and cell area (Y-axis) relative to the positive control HDAC inhibitor, TSA; % inhibition by TSA was set to 100% for each plate. Reduction in the number of identified cells (nuclei) compared to the positive control TSA was used as an indicator of toxicity. Toxic compounds are indicated in red, and were removed from subsequent analysis. (B) Class I hits are those compounds that significantly reduce cell area and ANF expression. Class II hits are compounds that significantly reduce cell area, but increase ANF expression.

J Mol Cell Cardiol, 2016, 97:106-13.. purchased from Selleck

Screening small molecule library in 384-well plates. (A): Schematic diagram of HTS screening of small molecule library. (B): Screening results of an epigenetics library. Each heat map represents one 384-well plate treated with the epigenetics library in duplicate wells at specified concentration(0.1, 1 and 10 μM). Wells G11 and K11 were duplicate wells of 5-aza-C. Wells K7 and L7 were duplicate wells of 5-aza-dC.

Stem Cells, 2017, 35(1):158-169.. purchased from Selleck

(c) Percentage of H3.3S31ph positive cells after treatment with candidate compound hits identified from the inhibitor library screen. The targets of inhibitors are the following: WZ3146: EGFR; Sorafenib Tosylate: VEGFR, PDGFR, and RAF/MEK/ERK; AT7519: CDK1, 2, 4, 5, 6, and 9; Hesperadin, AZD1152-HQPA, and ZM447439: AURKB; AZD8055 and KU-0063794: mTOR; MK-2206: Akt1, 2, and 3; GSK2126458: PI3K and mTOR.

J Mol Biol, 2017. . purchased from Selleck

Plots of locomotor seizure behaviour for 5 dpf scn1Lab mutants screened against (A) 52 ion channel ligands. Threshold for inhibition of seizure activity (positive hits) was determined as a reduction in mean swim velocity of 540% (red line). Blue data points represent compounds that were classified as toxic as treated larvae have no visible heartbeat or movement in response to touch after 90-min exposure.

Brain, 2017.. purchased from Selleck

Plots of locomotor seizure behaviour for 5 dpf scn1Lab mutants screened against (B) 254 compound GPCR ligands. Threshold for inhibition of seizure activity (positive hits) was determined as a reduction in mean swim velocity of 540% (red line). Blue data points represent compounds that were classified as toxic as treated larvae have no visible heartbeat or movement in response to touch after 90-min exposure.

Brain, 2017.. purchased from Selleck

The screening of Aβ40 inhibitors using AD models.(a) Scheme showing the cell culture procedure of chemical screening. (b) The result of first screening of Aβ40 inhibitors. The red line shows the value of the average ratio minus double the standard deviation (Ave - 2SD = 0.33). The compounds that reduced the Aβ40 ratio below Ave - 2SD value (0.33) passed the first screening. The Aβ40 level of DMSO treatment was considered to be 1.0. (c) The result of second screening in Aβ40 inhibitors. The red line indicates the criterion of hit compounds in the second screening (=0.5). (d,e) The effects of Aβ inhibition (d) and cell survival (e) in dose-response experiments using K1-derived neurons. These results show that PS1-overexpression did not influence the screening of Aβ inhibitors. The numerical data is shown in Supplementary Table S2. The amount of Aβ40 and cell viability in DMSO-treated PS1-G378E or K1 neurons was defined as 1.0. *P < 0.05, as determined by Steel’s test. Three independent experiments, each time in triplicates were performed (n = 3). Values are Mean ± SD. AraC, cytosine arabinoside; G378E, PS1-G378E neurons; K1, KhES-1-derived neurons; Nilo, Nilotinib; Pime, Pimecrolimus; Rosu, Rosuvastatin Calcium; Sulc, Sulconazole Nitrate salt; Tore, Toremifene Base.

Sci Rep, 2016, 6:33427.. purchased from Selleck

(A)Schematic view of virtual screening workflow on human ABCB1 and human ABCG2. (B) The sensitization effects of virtual screened compounds on SW620/Ad300 towards doxorubicin. (C) The sensitization effects of virtual screened compounds on NCI-H460/MX20 cells towards mitoxantrone. (D) Chemical structure of bafetinib.

Sci Rep, 2016, 6:25694.. purchased from Selleck

The results of the high-throughput screening (HTS) of 1017 FDA-approved drugs in the DSP assay for MERS-S. Screening results of 1017 drugs using the HTS DSP assay. (A) The vertical axis shows the reading of the DSP activity (RL activity) for the tested drugs (1 μM). The RL values were normalized to the control, which contained DMSO only. The horizontal axis represents the identification number arbitrarily assigned to each drug. Each dot represents an individual drug. The dotted line indicates 20% of the control. The value of the most active drug, nafamostat, is indicated (1.66% of the control).

Antimicrob Agents Chemother, 2016, 60(11):6532-6539.. purchased from Selleck

Relative wound density and cell viability z-score data of the shortlisted candidates from the primary screening assay. Based on their effect on migration and cell viability, the drugs were binned into high confidence and medium confidence migration and cytotoxic classes. a The migration z-score of the candidates are shown, drugs with a z-score between the dotted line (-1.6) and the straight line (–2) were binned into medium confidence migration class and the drugs with z-scores beyond the straight line (<-2) were binned as.

Clin Exp Metastasis, 2016, 33(4):385-99.. purchased from Selleck

Screen of library of FDA-approved library validates the adenylate kinase assay and indicates it is more sensitive than growth-based assays. A. Scatter plot of raw data from primary screen of the Selleck FDA-approved drug library. The cut-off for hit identification (2-fold increase in AK activity) is indicated by the solid line.

PLoS One, 2015, 10(6):e0129234.. purchased from Selleck

355 compounds from the Selleck library were docked into the six generated PRK1 homology models, the obtained docking poses were minimized using an implicit solvent model, and BFE calculation was subsequently performed. The predicted pIC50 was calculated using the developed QSAR model.

ChemMedChem, 2016, 11:1-12.. purchased from Selleck

Chemical screen targeting erythroid enucleation. (B) For the compound screen orthochromatic erythroblasts were isolated by FACS and subsequently incubated in 96-well plates in the presence of the compounds for 5h. The extent of enucleation was assessed by FACS LSR II. Graphs showing that the raw data between the duplicate assay plates are reproducible. (C) Graph showing compounds confirmed to significantly (paired student's t-test) inhibit enucleation compared to the vehicle control (DMSO). Data are means (+/− SD) of 4 independent experiments. *P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001 (paired student's t-test).

purchased from Selleck

Anti-ferroptosis activity of natural product. PANC1 cells were treated with erastin (20 mM) in the absence or presence of the indicated natural product compounds (10 mM). Cell viability was assayed using the CCK-8 kit

Biochem Biophys Res Commun, 2016, 473(4):775-80.. purchased from Selleck

Discovery of small-molecule inhibitors of MST3 using DSF. a) Distribution of compounds as a function of △Tm values. Of the 277 compounds screened, 23 compounds interfered with the assay due to intrinsic fluorescence or precipitation and were discarded from further evaluation; 109 compounds showed negative temperature shifts (-2.8 to -0.01℃) and were denoted as zero. Each bar represents compounds with △Tm ≤x-axis value, that is, 0: △Tm≤‹0; 1: △Tm≤1, etc. b) Identifiers and associated IC50 and △Tm values of compounds confirmed as MST3 binders by X-ray crystallography. c) Logarithmic proportionality between DTm and IC50 values; data were fit to Equation (1), yielding y0=4.08œ±0.3 and a=-0.93œ±0.09.

ChemMedChem, 2016, 11(11):1137-44.. purchased from Selleck

Hits of high throughput in vitro drug screen of three MLS cell lines. List of 27 drugs with a reduction in cell viability of >50% in two or all three cell lines at a drug concentration of 100 nM. Strong inhibitory effect of the survivin inhibitor YM155 is observed in two out of three MLS cell lines. Also, a good response is observed to several conventional chemotherapeutics, like doxorubicin, gemcitabine, and paclitaxel. Per drug, four concentrations (1, 10, 100, and 1000 nM) are tested; green boxes correspond to a high cell viability (~100%) and red boxes to a loss of cell viability (~0%). For comparison, at the bottom, five compounds are randomly shown that did not meet the criteria.

Transl Oncol, 2017, 10(4):546-554. purchased from Selleck

CCRF-CEM and Jurkat cells display a differential response to kinase inhibitors: Cells were treated with 100 nM (A) and 1000 nM (B) concentration of a panel of kinase inhibitors (378 inhibitors) for two days. Cell viability was measured using PrestoBlue cell viability assay. Selective inhibitors from “A” and “B” were used in figure "C" and "D".

Cancer Lett, 2017, 405:73-78. purchased from Selleck

The impact of FDA-approved drugs on ERa levels in MCF-7 cells. (A) Pie diagrams depicting the pharmaceutical categories in the library. (B) Synthetic protocol used for drug administration to MCF-7 cells. (C) Assay data depicting the quantitation of ERa levels in each one of the 162 performed Western blots. Z0 is the Z factor for negative (DMSO) and positive (E2) controls in each Western blot analysis. Robust Z scores (Z*) graphs for drug-treated samples alone (D), in combination with E2 (E) or for the effect of E2 within each drug-treated sample (i.e., E2þdrug/drug alone) (F); red arrows indicate the Z* for drugs considered as positive hits. Red lines indicate the threshold used for analysis. Western blot analyses of ERa and cathepsin D (Cat D) expression levels in MCF-7 cells treated for 24 h with E2 (1 nM) both in the presence or in the absence of 100 nM estriol (ESTR) (G), coumarin (COUM) (G0), basedoxifene (BASE) (H), vinblastine (VINB) (H0) and carfilzomib (CARF) (I) or vehicle (DMSO-CTR). The loading control was done by evaluating vinculin expression in the same filter.

Mol Cell Endocrinol, 2017, S0303-7207(17)30400-8. purchased from Selleck

(A) Schematic representation of the compound screening. J-LAT A2 cells were co-treated with compounds (5 μM) and TNFα (10 ng/ml) for 24 hr. Cells were then stained with Hoechst® 33342 followed by imaging analysis. The assay was performed in quadruplicate. (B) Fluorescence images showing the reactivation (GFP, green) and cell nucleus (blue) in J-LAT A2 cells treated with TNFα for the indicated compounds. Values represent the mean ± s.d. of the % GFP-positive cells (n ≥ 4). (C) Scatter plot of the % GFP-positive cells vs % cell numbers for all tested compounds. Results were normalized to DMSO control. LSM, levosimendan; SPR, spironolactone; 9AA, 9-aminoacridine

Antiviral Res, 2017, 146:76-85. purchased from Selleck

High-throughput luciferase screening to identify parkin inducing compounds. (A) Schematic illustration of the high-throughput screening method. HEK-293T parkin reporter cell line (parkin-LucHEK-293T) was selected by stable transfection of the luciferase construct containing three repeats of parkin promoter’s CREB/ATF4 binding motifs. In a 96-well plate, 1172 FDA-approved drugs were used to treat parkinLuc-HEK-293T cells. Parkin promoter activity was measured by luciferase assay. DMSO was used as a negative control. CCCP treatment was used as a positive control. (B) Parkin promoter activities in parkin-Luc-HEK-293T cell line treated with DMSO or 10uM CCCP as positive control (n=4) were measured by luciferase assay. (C) Scatter plot showing relative increase of parkin promoter activity in parkin-Luc-HEK-293T cell line treated with each compound compared to DMSO negative control based on luciferase assay. Top 20 compounds were highlighted in red color. (D) Relative increase of parkin promoter activity induced by the top 20 compounds based on the initial HTS screening determined by luciferase assay (n=9, see also Table 1). Quantified data are expressed as mean±s.e.m. *P<0.05, **P<0.01, and ***P<0.001, nonparametric Kruskal-Wallis ANOVA test (B) and ANOVA test followed by Tukey post hoc analysis (D).

Sci Rep, 2017, 7(1):525. purchased from Selleck

(c) Results of the second-round test on Aurora kinases inhibitors. Samples are under corresponding treatment for 3 weeks and measured by bioluminescence. CTRL, control. P value is determined by two-tailed unpaired Mann-Whitney U-test with multiple-test correction. N=6 bone fragments in each group. Representative bioluminescence images are shown at the bottom of the panel. (d) Results of the second-round test on HMT inhibitors. Samples are under corresponding treatment for 3 weeks and measured by bioluminescence. P value is determined by two-tailed unpaired Mann–Whitney U-test with multiple-test correction. N=6 bone fragments in each group. Representative bioluminescence images are shown at the bottom of the panel.

Nat Commun, 2017, 8:15045. purchased from Selleck

Screening small molecule library in 384-well plates. (A) Schematic diagram of HTS screening of small molecule library. (B) Screening results of an epigenetics library (128 compounds; see Table S1 for a description of compounds in each well). Each heat map represents one 384-well plate treated with the epigenetics library in duplicate wells at specified concentration (0.1, 1 and 10 μM). Wells G11 and K11 were duplicate wells of 5-aza-C. Wells K7 and L7 were duplicate wells of 5-aza-dC.

Stem Cells, 2017, 35(1):158-169. purchased from Selleck

Emodin was found to inhibit DPP4 activity after screening a natural compound library.

PeerJ, 2017, 5:e3283. purchased from Selleck

c Heat map of selected anti-cancer compounds exhibiting strong inhibition in at least one of the PDC lines. The scale represents percentage inhibition of the compounds, with inhibition score <50% shown in grey. d Selected molecular signatures (P < 0.05) of genes that show elevated expressions across the five Met cell lines, some of which appear to be associated with the selective responses of PDC lines to compounds of same target classes.

Nat Commun. 2017, 8(1):435. purchased from Selleck

Example of screen results for a pair of single 384-well plates treated with drug library only (top) and drug library plus 400 nM MK1775 (bottom). The histograms show counts versus γH2AX median values in S-phase.

Oncotarget, 2017, 8(7):10966-10979. purchased from Selleck

Repurposing Library contents. (a) Highest clinical phase achieved by each compound. 3,422 drugs in the library have reached clinical use as of November 2016. (b) Number of compounds per target category. Compounds with multiple targets might be indicated in more than one category. (c) Number of approved drugs for indications within each listed disease area. (d) Classification of proteins targeted by library drugs. Protein function hierarchy is shown with increasing specificity of function corresponding to distance from the figure center. The relative area of each segment is proportional to the fraction of the Repurposing Library targeting each protein class. As expected, the library is enriched in drugs targeting kinases, GPCRs, and ion channels.

Nat Med, 2017, 23(4):405-408. purchased from Selleck

scatter plot representing percent growth inhibition of WT L. monocytogenes in the presence of a combination of a sublethal dose (1 μg/ml) of the β-lactam ceftriaxone and each compound in the screen. The solid black line represents the library mean (μ), and the dashed black line and gray line represent two (2σ) and three (3σ) S.D. above the library mean, respectively. The cyan, green, and red data points represent staurosporine, GSK690693, and other compounds from the IPA family, respectively.

J Biol Chem, 2017, 292(41):17037-17045. purchased from Selleck

(A) HTS assay timeline. Vero cells were seeded at a density of 1 × 10⁴ cells per well in 96-well plates. After overnight incubation, cells were treated in duplicate with 10 μM compounds. One hour later, 20 μl RVPs was added. The supernatant was removed 1 h later and the compounds were readded to the cells for an additional 23 h. (B) (Left) HTS assay flowchart. The criterion required for the compounds to pass the primary screen was inhibition of >90%, and 28 primary candidates were selected. In the reconfirmation screen, 9 compounds with dose-dependent inhibition and cell viability of >80% were selected. (Middle) HTS of a library of 1,018 FDA-approved drugs for primary candidates inhibiting JEV infection. Each dot represents the percent inhibition achieved with each compound at a concentration of 10 μM. The dots located in the blue box represent inhibition of >90%. (Right) IC50s, CC50s, and SIs of nine compounds selected from the reconfirmation screen. The top five compounds depicted in the blue box showed SIs of greater than 10 and were designated hit drugs.

J Virol, 2017, e01055-17. purchased from Selleck

Normalized effect of inhibitors grouped according to their primary targets on SmoIP fluorescence. Effect of Hh stimulation on untreated cells set as 100% (dashed line). Diameter of circle reflects number of components per cluster, cutoff N ≥ 3, clusters sorted along X axis accordingly. Inhibitors used at 15 μM.

Sci Rep, 2017, 7(1):14341. purchased from Selleck

Select statin class drugs lysed S. mutans in an AK assay screen of the Selleck library. Statins were identified in the screening of the Selleck library against planktonic cultures of S. mutans UA159. The data are representative of 2 independent screens.

Antimicrob Agents Chemother, 2017, e01674-17. purchased from Selleck

Effects of compound 18n (A,B) and compound 20b (C,D) on the ROS of of HepG2 cells. The level of intracellular reactive oxygen species (ROS) in HepG2 cells was measured by flow cytometry/ Data were shown as mean ± SEM (n = 3).

Eur J Med Chem, 2017, 140:421-434. purchased from Selleck

High-content drug screening. A, Schematic of the screening process and hit selection. Phases of 3D screening and follow-up drugs testing are listed on the left. Numbers represent the considered candidate compounds after each phase. B, Examples of brightfield and fluorescent images of DMSO-treated MCS obtained from three NB cell lines. C, Illustration of image analysis with an in-house–developed algorithm evaluating spheroids viability. Values in black correspond to the spheroid area as compared with vehicle-treated controls, whereas the ratio of calcein fluorescence in the rim and center of each spheroid is in red. Zero multiplier (light green) is introduced when calcein staining of a spheroid is comparable with the background. D, Scatter plot demonstrating the results of the primary screening in CHP-134. Each dot represents one compound, and the white color indicates primary hits. E, Venn diagram showing the number of primary hits (n ¼ 193), among which 60 are FDA-approved, emerged as potentially effective in at least two NB cell lines.

Mol Cancer Ther, 2018, doi: 10.1158/1535-7163.MCT-17-0841. purchased from Selleck

High-throughput luciferase screening to identify parkin inducing compounds. (A) Schematic illustration of the high-throughput screening method. HEK-293T parkin reporter cell line (parkin-Luc-HEK-293T) was selected by stable transfection of the luciferase construct containing three repeats of parkin promoter's CREB/ATF4 binding motifs. In a 96-well plate, 1172 FDA-approved drugs were used to treat parkin Luc-HEK-293T cells. Parkin promoter activity was measured by luciferase assay. DMSO was used as a negative control. CCCP treatment was used as a positive control. (B) Parkin promoter activities in parkin-Luc-HEK-293T cell line treated with DMSO or 10uM CCCP as positive control (n=4) were measured by luciferase assay. (C) Scatter plot showing relative increase of parkin promoter activity in parkin-Luc-HEK-293T cell line treated with each compound compared to DMSO negative control based on luciferase assay. Top 20 compounds were highlighted in red color. (D) Relative increase of parkin promoter activity induced by the top 20 compounds based on the initial HTS screening determined by luciferase assay (n=9, see also Table 1). Quantified data are expressed as mean±s.e.m. *P<0.05, **P<0.01, and ***P<0.001, nonparametric Kruskal-Wallis ANOVA test (B) and ANOVA test followed by Tukey post hoc analysis (D).

Sci Rep, 2017, 7(1):525. purchased from Selleck

Compound 1773 inhibition of the enzymatic activity of recombinant Legumain. (A) 10 μM of compound 1773 was incubated with recombinant Legumain and the fluorogenic substrate. (B) Specificity testing of compound 1773. 1 or 10 μM of compound 1773 was incubated with different concentrations of AMC (0–64 μM) alone and fluorescence measured every 2 min for 20 cycles. Compound 1773 was screened out from 500 compounds library (FDA-Approved-Drug-Library, Selleck, USA).

Protein Expr Purif, 2018, 150:12-16. purchased from Selleck

(c) Percentage of H3.3S31ph positive cells after treatment with candidate compound hits identified from the inhibitor library screen. The targets of inhibitors are the following: WZ3146: EGFR; Sorafenib Tosylate: VEGFR, PDGFR, and RAF/MEK/ERK; AT7519: CDK1, 2, 4, 5, 6, and 9; Hesperadin, AZD1152-HQPA, and ZM447439: AURKB; AZD8055 and KU-0063794: mTOR; MK-2206: Akt1, 2, and 3; GSK2126458: PI3K and mTOR.

J Mol Biol, 2017, 429(13):2042-2045. purchased from Selleck

Phosphorylation of CREB affected by different concentrations of TG100-115 determined by using immunoblotting. TG100-115 was screened out from a kinase inhibitors library for highly potent inhibiting TRPM7 kinase.

Biochim Biophys Acta, 2017, 1861(4):947-957. purchased from Selleck

LeX-specific antibody SSEA-1 was preincubated with different concentrations (1–80 μM) of gossypol (red), folic acid (blue), orlistat (orange), tosufloxacin tosylate (purple), ursolic acid (green), or the control compound idarubicin (black) and added to wells coated with the LeX glycomimetic peptide. The signal of LeX-specific SSEA-1 antibody alone without any competing chemical compound was set on 100%. Asterisks signify differences in competition between all LeX mimetic compounds and the negative control compound idarubicin as calculated by one-way analysis of variance (ANOVA) followed by Fisher’s PLSD test (F = 9.468, p = 0.0001; PLSD *p < 0.05). Gossypol and ursolic acid were from the Natural Product Library.

Mol Neurobiol, 2018, doi: 10.1007/s12035-018-0953-8. purchased from Selleck

PC9GR, H1650, and H1975 cells were treated with 124 compounds in a natural product library. These compounds showed as more than 50% cytotoxicity of these three cell types.

Int J Biol Sci, 2018, 14(1):47-56. purchased from Selleck

In vitro anti-fibrotic drug screening for the FDA-approved drugs 1-75 (a) and FDA-approved drugs 76-150 (b).

J Cell Physiol, 2018, 233(1):422-433. purchased from Selleck

HEK293T cells were transfected with luciferase reporter driven by NF-jB response elements for 24 h, and treated with 104 drugs or DMSO for 12 h. Luciferase activity was determined using the Dual-Luciferase Reporter Assay System.

Cancer Sci, 2018, 109(4):1220-1229. purchased from Selleck