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3-Methyladenine (3-MA) Autophagy/PI3K inhibitor

Name: 3-Methyladenine (3-MA)
Brand: Selleck Chemicals
SKU: S2767
Availability: InStock
Rating: 5 (956 reviews)

Cat.No.S2767

3-Methyladenine (3-MA) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells. It blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also inhibits autophagosome formation. Solutions are unstable and should be fresh-prepared.

Chemical Structure

Molecular Weight: 149.15

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Quality Control

Batch: Purity: 99.97%

99.97

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Related Targets	Akt mTOR GSK-3 ATM/ATR DNA-PK AMPK PDPK1 PTEN PP2A PDK
Other PI3K Inhibitors	GDC-0077 (Inavolisib) SAR405 Quercetin (Sophoretin) LY294002 XL147 analogue Tersolisib (STX-478) Buparlisib (BKM120) 740 Y-P (PDGFR 740Y-P) GO-203 TFA Eganelisib (IPI-549)

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
K562	Function Assay	10mM	1h		decreases the expression of LC3-II and the formation of autophagosomes	21864037
Jurkat	Function Assay	10mM	1h		decreases the expression of LC3-II and the formation of autophagosomes	21864037
HeLa	Cytotoxicity Assay	2mM	24h		inhibites the cytotoxicity of silibinin to HeLa cells.	21875385
PC12/TetOn	Function Assay	0.1/1mM	18h		leads to α-syn(WT) accumulation, toxicity, and oligomer formation	21906659
RMPI8226	Function Assay	5mM	1h		suppresses the level of autophagy under nutrient depletion	21915620
MCF-7	Function Assay	10mM	48h		blocks autophagy induced by bortezomib	21931937
HBx	Apoptosis Assay	10mM	48h	DMSO	increases cell death	22020078
Marc-145	Function Assay	5mM	12/24/36h		reduces the PRRSV titers and the protein expression	22119900
U937	Function Assay	2mM	12h		decreases the autophagy ratio	22155150
BGC-823	Function Assay	5mM	2h		inhibits the rate of autophagic cells	22322152
A549	Function Assay	0.1mM	24h		suppresses SU11274-induced cell death	22466960
pDCs	Function Assay	10mM	0.5h		reduces the induction of IFN-α by ssRNA40	22396599
HeLa	Apoptosis Assay	5mM	24h		induces caspase-dependent cell death	22545128
U251	Apoptosis Assay	5mM	24h		increases S1-induced cell death	22579788
MCF-7	Apoptosis Assay	0.1mM	6h		enhances sirtinol-induced apoptosis	22751989
PC-3	Apoptosis Assay	2mM	2h		increases ORI-induced cell death	22745580
HCT116	Apoptosis Assay	5mM	24h	DMSO	enhances apigenin-induced cell death	24626522
U2OS	Growth Inhibition Assay	10mM	24h		intensifies the growth inhibition induced by Dox	24639013
A2780cp	Apoptosis Assay	2.5mM	1h	ddH2O	enhances cisplatin-induced cell death	24817946
HepG2	Function Assay	5mM	4h		increases cellular levels of HL	24713587
Microglia	Apoptosis Assay	5mM	24h		decreases hypoxia-induced cell death	24818601
MDA-MB 231	Apoptosis Assay	5mM	0.5h		modulates Tocomin® induced apoptosis	24830781
PANC-1	Apoptosis Assay	1mM	48h	DMSO	enhances bortezomib-induced cell viability loss	24842158
MDA-MB-231	Function Assay	2mM	48h		promotes TM-induced cell death	24970676
MDA-MB-231	Function Assay	2mM	24h		inhibits autophagy induced by TM	24970676
MCF-7	Function Assay	2mM	48h		promotes TM-induced cell death	24970676
MCF-7	Function Assay	2mM	24h		inhibits autophagy induced by TM	24970676
HepG2	Apoptosis Assay	3mM	5h		reduces cell apoptosis induced by QDs	22836595
HeLa	Apoptosis Assay	10mM	2h		decreases cell viability co-treatment with PEI	23000135
SK-HEP-1	Apoptosis Assay	10mM	1h		protects against autophagy and induces apoptosis in bufalin-treated cells	22858649
MDA-MB231	Function Assay	5mM	1h		increases resveratrol-mediated caspase activation and cell death	23088850
PaCa44	Apoptosis Assay	2.5mM	1h		reduces genipin-mediated apoptosis	23124112
T-47D	Function Assay	10mM	2h		inhibits autophagy process and increases rapamycin induced apoptosis	23300026
GTL-16	Apoptosis Assay	5mM	24h		reduces cell viability as compared to cells treated with MET inhibitors	23313490
U251MG	Function Assay	3mM	1h		suppresses LC3-II protein expression	23338618
T24	Function Assay	10mM	1h		reduces the cleavage of LC3 after baicalin treatment	23354080
HUVECs	Function Assay	3mM	24h		blocks the protective effect of resveratrol by inhibiting autophagy	23358928
MCF-7	Function Assay	5mM	24h		inhibits starvation-induced autophagy	23395679
Hela	Function Assay	5mM	24h		inhibits starvation-induced autophagy	23395679
OR6	Function Assay	10mM	72h		suppresses HCV replication and formation of autophagosomes	23395875
HT-29	Function Assay	1mM	48/96h		inhibits AMPK induces autophagic cell death	23508272
SH-SY5Y	Cytotoxicity Assay	5mM	24h		increases PCN toxicity	23525265
Saos-2	Apoptosis Assay	1mM	96h		increases cell death induced by PCX	23563171
1321N1	Cytotoxicity Assay	5mM	24h		protects cell against PCN-induced toxicity	23525265
A2780	Apoptosis Assay	5mM	24h		converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells	23592281
OV2008	Apoptosis Assay	5mM	24h		converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells	23592281
PC12	Function Assay	10mM	24h	water	inhibits chymotrypsin-like proteasomal activity.	23603979
SH-SY5Y	Apoptosis Assay	5mM	1h		abolishes celastrol neuroprotective effect	23619395
SH-SY5Y	Function Assay	1mM	24h		inhibits the autophagy induced by TOCP	23743148
HepG2	Function Assay	10mM	24h		inhibits siTIGAR- and HBSS-induced autophagy	23817040
HeLa	Function Assay	10mM	2h		suppresses LC3 II expressison	23864738
HONE-1	Function Assay	5mM	1h		represses 6r-mediated ROS production	23892358
MCF7	Function Assay	5mM	24h		increases CuO induced cell death	23962629
HO8910	Apoptosis Assay	10mM	12h		enhances B19-induced apoptosi	23983610
SMMC-7721	Apoptosis Assay	5mM	24h		attenuates TNF-α protection against serum starvation-mediated apoptosis	24066693
Hep3B	Apoptosis Assay	5mM	24h		attenuates TNF-α protection against serum starvation-mediated apoptosis	24066693
H460	Function Assay	10mM	4h		increases cisplatin-induced cell death	24173208
A549	Function Assay	10mM	4h		inhibits autophagy induced by irradiation	24142735
H1299	Function Assay	10mM	4h		increases cisplatin-induced cell death	24173208
WiDr	Function Assay	10mM	1h		inhibits PCBL-induced LC3 II expression	24190489
LoVo	Apoptosis Assay	5mM	48h		enhances DCA-induced apoptosis.	24201812
HepG2 E47	Function Assay	2.5mM	48h		increases the toxicity of AA, BSO, and CCl4	24273738
RKO	Function Assay	2mM	1h	DMSO	enhances cell death by geldanamycin	24291777
Hep3B	Apoptosis Assay	2mM	12h	DMSO	inhibits AZD8055-induced cell death	24297300
ACHN-5968	Apoptosis Assay	5mM	3h		enhances paclitaxel-mediated apoptosis	24305604
Huh7	Apoptosis Assay	2mM	12h	DMSO	inhibits AZD8055-induced cell death	24297300
UOK257	Apoptosis Assay	5mM	3h		enhances paclitaxel-mediated apoptosis	24305604
ECSCs	Apoptosis Assay	10mM	4h		decreases rapamycin-treated apoptosis	24319109
MCF-7	Function Assay	10mM	24h		inhibits the autophagy induced by chemotherapy drugs	24315578
SGC-7901	Apoptosis Assay	2mM	1h		increases CA-4 induced apoptosis	24321340
SMMC-7721	Apoptosis Assay	2mM	1h		increases CA-4 induced apoptosis	24321340
T24	Apoptosis Assay	5mM	1.5h		potentiates celecoxib-induced apoptosis	24349176
NTUB1	Apoptosis Assay	5mM	1.5h		potentiates celecoxib-induced apoptosis	24349176
MG-63	Apoptosis Assay	10mM	12h		enhances DP-induced apoptosis	24358301
MG-63	Apoptosis Assay	0.5/1mM	32h		enhances salinomycin-induced cell apoptosis	24358342
MG-63	Function Assay	0.5/1mM	48h		induces salinomycin-induced cell viability loss	24358342
U2OS	Function Assay	0.5/1mM	48h		induces salinomycin-induced cell viability loss	24358342
HGC-27	Function Assay	10mM	1h		inhibits the cell viability loss by RAD001 or MK-2206	24416349
HCT116	Apoptosis Assay	5mM	24h		enhances the apoptosis induced by apigenin	24626522
A549	Apoptosis Assay	10mM	48h		accelerates the reduction of cell viability induced by PTX	24626722
Saos-2	Apoptosis Assay	10mM	24h		intensifies the growth inhibition of the U2OS cells induced by Dox	24639013
U2OS	Apoptosis Assay	10mM	24h		intensifies the growth inhibition of the U2OS cells induced by Dox	24639013
HepG2	Function Assay	5mM	4h		increases HL release	24713587
A549	Apoptosis Assay	5mM	48h		decreases the percentage of cell death and expression levels of caspase-3, Beclin-1 and LC3-II	24706303
A2780cp	Apoptosis Assay	2.5mM	1h	ddH2O	enhances cisplatin-induced cell death	24817946
Microglia	Apoptosis Assay	5mM	24h		decreases hypoxia-induced cell death	24818601
HT-29	Apoptosis Assay	1mM	48h	DMSO	enhances bortezomib-induced cell viability loss	24842158
MDR	Apoptosis Assay	10mM	6h		strengthens the power of anticancer drugs	25019701
H157	Function Assay	5mM	2h		suppresses SPC induced accumulation of LC3-II	25285628
A549	Function Assay	5mM	2h		suppresses SPC induced accumulation of LC3-II	25285628
A2780cp	Growth Inhibition Assay	1mM	1h		increases cisplatin-induced cell death	25322694
NBL-W-S	Apoptosis Assay	1mM	6h		increases cell apoptosis induced by GANT-61	25323222
NBL-W-S	Growth Inhibition Assay	1mM	6h		enhances GANT-61 toxicity	25323222
A549	Apoptosis Assay	5mM	2h	DMSO	inhibits BDMC-induced apoptotic cell death	25716561
95D	Apoptosis Assay	5mM	2h	DMSO	inhibits BDMC-induced apoptotic cell death	25716561
A549	Growth Inhibition Assay	3mM	2h	DMSO	reduces growth inhibitory effect of BDMC	25716561
95D	Growth Inhibition Assay	3mM	2h	DMSO	reduces growth inhibitory effect of BDMC	25716561
Nara-H	Growth Inhibition Assay	5mM	48h		enhances temsirolimusmediated suppression of Nara-H cell proliferation	21805033
HUVECs	Function Assay	10mM	0.5h		decreases the AGE-BSAinduced autophagy leve	21468592
HepG2	Apoptosis Assay	2mM	1h		enhances radiation-induced cell death	21453691
U-2 OS	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells	29435139
A673	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
DAOY	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells	29435139
Saos-2	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
BT-37	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
RD	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
SK-N-SH	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
BT-12	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells	29435139
MG 63 (6-TG R)	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells	29435139
NB1643	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
Rh41	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
Rh30	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh30 cells	29435139
SJ-GBM2	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
SK-N-MC	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB-EBc1	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
LAN-5	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
Rh18	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells	29435139
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 10 mg/mL (67.04 mM) Warmed with 50°C water bath; Ultrasonicated;
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 10 mg/mL

Water : 4 mg/mL

DMSO : 16 mg/mL (107.27 mM) Warmed with 50°C water bath; Ultrasonicated;
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : 10 mg/mL (超声加热十分钟，60℃)

Ethanol : 8 mg/mL

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.750mg/ml (5.03mM)	Taking the 1 mL working solution as an example, add 50 μL of 15 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.500mg/ml (3.35mM)	Taking the 1 mL working solution as an example, add 50 μL of 10 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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Average weight of animals: g

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% DMSO

% Tween 80

% ddH₂O

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Chemical Information, Storage & Stability

Molecular Weight	149.15	Formula	C₆H₇N₅	Storage (From the date of receipt)	3 years -20°C powder
CAS No.	5142-23-4	Download SDF		Storage of Stock Solutions	Solutions are unstable. Prepare fresh or purchase small, pre-packaged sizes. Repackage upon receipt.
Synonyms	NSC 66389			Smiles	CN1C=NC(=N)C2=C1N=CN2

Mechanism of Action

Targets/IC₅₀/Ki	Autophagy Vps34 (HeLa cells) 25 μM PI3Kγ (HeLa cells) 60 μM
In vitro	The slight preference for Vps34 prevention by 3-Methyladenine (3-MA) probably arises from a hydrophobic ring specific to Vps34, which encircles the 3-methyl group of this compound. It has been reported to cause cancer cell death under both normal and starvation conditions, and could also suppress cell migration and invasion independently of its ability to inhibit autophagy, implying that it possesses functions other than autophagy suppression. This compound elicits caspase-dependent cell death that is independent of autophagy inhibition. Treatment with 5 mM of it reduces the percentage of glucose-starved HeLa cells displaying GFP-LC3 puncta to 23%. The levels of LC3-I are increasing and the levels of LC3-II are decreasing between 12 and 48 hours in cells that are treated with 3-MA. Conversion of LC3-I to LC3-II is suppressed by the compound. Treatment of HeLa cells with it at 2.5 mM or 5 mM for one day does not affect cell viability, whereas treatment with 10 mM for one day causes a 25.0% decrease in cell viability. Treatment of cells with 2.5, 5 or 10 mM for two days causes 11.5%, 38.0% and 79.4% decrease in viability, respectively. It decreases cell viability in a time- and dose-dependent manner, and significantly shortens the duration of nocodazole-induced-prometaphase arrest. Suppression of autophagy by 3-MA inhibits SU11274-induced cell death. Prolonged treatment with it (up to 9 hours) induces significant LC3 I to II conversion in wild type MEFs. Prolonged treatment with 3-MA, but not wortmannin, markedly increases GFP-LC3 punctuation/aggregation. Its induced LC3 conversion and free GFP liberation are ATG7-dependent. Treatment with it leads to evident increase of p62 protein level. The compound increases the p62 level even in Atg5−/− MEFs as well as in cells with DOX-mediated deletion of ATG5. It inhibits class I and class III PI3K in different temporal patterns. Its induced LC3 I to LC3 II conversion is dramatically compromised in Tsc2−/− cells compared with wild type cells. This compound disrupts the anti-autophagic function of mTOR complex 1.
Kinase Assay	Protein degradation assay
Kinase Assay	HeLa cells are radiolabeled for 24 hours with 0.05 mCi/mL l-[U-¹⁴C]valine. At the end of the labeling period, cells are rinsed three times with PBS. Cells are incubated for the designated times in either full medium or EBSS with or without the presence of 10 mM 3-Methyladenine (3-MA).
In vivo	3-Methyladenine (3-MA) blocks autophagy through its effect on class III phosphatidylinositol 3-kinase (PI3K). Treatment with this compound does not alter the degree of hemorrhage compared with the subarachnoid hemorrhage (SAH) group. Its pretreatment significantly aggravates neurological symptoms when compared with the SAH + vehicle group. Autophagy is decreased when it is applied. Conversely, cleaved caspase-3 is markedly up-regulated in the SAH + 3-MA group. In line with the up-regulation of cleaved caspase-3 expression, the number of TUNEL-positive cells in the right cortex is significantly increased in the SAH + 3-MA group compared with the SAH + vehicle group.
References	[1] https://pubmed.ncbi.nlm.nih.gov/20574157/ [2] https://pubmed.ncbi.nlm.nih.gov/22545128/ [3] https://pubmed.ncbi.nlm.nih.gov/22466960/ [4] https://pubmed.ncbi.nlm.nih.gov/20123989/ [5] https://pubmed.ncbi.nlm.nih.gov/22521819/

Applications

Methods	Biomarkers	PMID
Western blot	α-SMA / TGF-β / LC-3BI / LC-3B II / Beclin-1 / NF-κB p65 caspase-3 / caspase-9 / PARP VEGF APP / BACE1 / ADAM17 / Presenilin 1 / Presenilin 2 / Nicastrin / APH-1 / Pen-2 / LC3-1 / LC3-2	29296191
Immunofluorescence	LC3 / Hif-α / COX2	29039446
Growth inhibition assay	Cell viability	26934124

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Handling Instructions

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Frequently Asked Questions

Question 1:
I'm also wondering whether it can be dissolved in water, or maybe something like culture medium, normal saline solution to form 10mM solution.

Answer:
As the reference (http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal. pone.0035665), it was found to inhibit autophagy at concentrations ranging from 1 to 10 mM and was directly dissolved into the culture medium at the indicated concentrations. And we tested the solubility of S2767, and found its solubility in DMEM is 31 mg/mL at about 40°C.

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