Antibiotics

Sensitivity of AML cells to conventional induction treatment and small-molecule p53 activators. Cell viability in OCI-AML3 cells treated for 24 hours with cytarabin/idarubicin (CI) and Nultin-3A (Nut; A), CI and Leptomycin-B (LMB; B), or Nut and LMB (D), respectively. CI was tested at 0, 100, 200, and 300 nmol/L CI; Nutlin-3A at 0, 2.5, 5, 7.5; and LMB at 0, 2, 8, 32 ng/mL in dosages 0, 1, 2, and 3, respectively. Cell viability in normal and AML bone marrow cells.

Cells were treated with brefeldin A or manumycin A, and the resulting supernatant was collected after 48 h for exosomal preparation (lanes 1 and 2), or exosomes obtained from C81 cells were trypsin-treated or freeze/thawed (F/T) and then trypsin-treated (lanes 3 and 4). Lanes 5 and 6, input exosome controls from C81 or CEM cells, respectively. Resulting exosomes were assayed for the presence of Tax by Western blotting.

Cell viabilities with increasing concentrations of cisplatin (CP) and doxorubicin (DOXO) under normoxic and hypoxic condition for 48 hours were determined by MTT assay. IC50 values are presented as the means ?SDs (n=4) and * denotes p<0.05.

Same experiment as in Figure 2 with fluorescent daptomycin shown as confocal images. A 7/3 DOPC/DOPG GUV (including 1% Rh-PE) was introduced at time zero into a solution containing 1 uM daptomycin with 0.5 uM BODIPY-daptomycin and 1 mM Ca2+. To show colocalization of lipid and peptide in aggregates, confocal images were taken: red for RH-PE and green for BODIPY-daptomycin. As the protrusion length decreased, aggregates appeared on the GUV surface containing both lipid and peptide. The scale bar is 10 um.

Effect of FDA approved drugs on EBOV transfected donor and recipient immune cells. (A) Varying concentrations of Cambinol and Esomepraole (1 nM, 1 μM, 10 μM), Oxytetracycline (1 nM, 10 nM, 10 μM), and DMSO (0.00001, 0.0001, 0.001%) treatments were administered to VP40-transfected 293T cells, followed by incubation for 3 days. Supernatant was centrifuged to remove cellular material and subsequently incubated for 72 h at 4°C with 20 μL of 30% slurry of NT80/82 beads. The NT pellet was washed twice with PBS and resuspended in SDS Laemmli buffer, followed by Western blot analysis for levels of exosomal markers CD63 and Alix. Actin levels were also analyzed. Mid-log phase EVTR2C cells (containing VP40) were treated with either low (0.1 μM), medium (1 μM), or high (10 μM) concentrations of Oxytetracycline and incubated for 5 days along with untreated 293T cells. Cell-free supernatants were harvested, filtered, and used to treat CEM (B), Jurkat (C), and U937 (D) cells. Cells were incubated for 5 days and assayed with CellTiter-Glo for viability. Statistical significance was determined using two-tailed student’s t-test with all groups compared to the untreated 293T cell control groups. RLU, Relative Luminescent Units. ∗p < 0.05, ∗∗∗p < 0.001.

Sensitivity of AML cells to conventional induction treatment and small-molecule p53 activators. Cell viability in OCI-AML3 cells treated for 24 hours with cytarabin/idarubicin (CI) and Nultin-3A (Nut; A), CI and Leptomycin-B (LMB; B), or Nut and LMB (D), respectively. CI was tested at 0, 100, 200, and 300 nmol/L CI; Nutlin-3A at 0, 2.5, 5, 7.5; and LMB at 0, 2, 8, 32 ng/mL in dosages 0, 1, 2, and 3, respectively. Cell viability in normal and AML bone marrow cells.

Cells were treated with brefeldin A or manumycin A, and the resulting supernatant was collected after 48 h for exosomal preparation (lanes 1 and 2), or exosomes obtained from C81 cells were trypsin-treated or freeze/thawed (F/T) and then trypsin-treated (lanes 3 and 4). Lanes 5 and 6, input exosome controls from C81 or CEM cells, respectively. Resulting exosomes were assayed for the presence of Tax by Western blotting.

Cell viabilities with increasing concentrations of cisplatin (CP) and doxorubicin (DOXO) under normoxic and hypoxic condition for 48 hours were determined by MTT assay. IC50 values are presented as the means ?SDs (n=4) and * denotes p<0.05.

Same experiment as in Figure 2 with fluorescent daptomycin shown as confocal images. A 7/3 DOPC/DOPG GUV (including 1% Rh-PE) was introduced at time zero into a solution containing 1 uM daptomycin with 0.5 uM BODIPY-daptomycin and 1 mM Ca2+. To show colocalization of lipid and peptide in aggregates, confocal images were taken: red for RH-PE and green for BODIPY-daptomycin. As the protrusion length decreased, aggregates appeared on the GUV surface containing both lipid and peptide. The scale bar is 10 um.

Effect of FDA approved drugs on EBOV transfected donor and recipient immune cells. (A) Varying concentrations of Cambinol and Esomepraole (1 nM, 1 μM, 10 μM), Oxytetracycline (1 nM, 10 nM, 10 μM), and DMSO (0.00001, 0.0001, 0.001%) treatments were administered to VP40-transfected 293T cells, followed by incubation for 3 days. Supernatant was centrifuged to remove cellular material and subsequently incubated for 72 h at 4°C with 20 μL of 30% slurry of NT80/82 beads. The NT pellet was washed twice with PBS and resuspended in SDS Laemmli buffer, followed by Western blot analysis for levels of exosomal markers CD63 and Alix. Actin levels were also analyzed. Mid-log phase EVTR2C cells (containing VP40) were treated with either low (0.1 μM), medium (1 μM), or high (10 μM) concentrations of Oxytetracycline and incubated for 5 days along with untreated 293T cells. Cell-free supernatants were harvested, filtered, and used to treat CEM (B), Jurkat (C), and U937 (D) cells. Cells were incubated for 5 days and assayed with CellTiter-Glo for viability. Statistical significance was determined using two-tailed student’s t-test with all groups compared to the untreated 293T cell control groups. RLU, Relative Luminescent Units. ∗p < 0.05, ∗∗∗p < 0.001.