Figure 4.
Venetoclax-gilteritinib combination reduces ERK and GSK3B phosphorylation and MCL-1 protein levels via proteasomal degradation. (A) Protein expression of MCL-1, BCL-2, pERK, and total ERK in HL60 cells treated for indicated timespan with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. B-actin levels are given as loading control. All western blot images have been cropped for improved clarity and conciseness. Quantification was performed using ImageJ. Data are representative of 3 independent experiments. (B) Protein expression of pGSK3A and B, total GSK3A and B, and MCL-1 in HL60 cells treated for indicated timespan with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. Vinculin levels are given as loading control. Quantification was performed using ImageJ. Data are representative of 3 independent experiments. (C) Protein expression of MCL-1, pMCL-1 S159, and pMCL-1 T163 in HL60 cells treated for indicated time points with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. B-actin levels are given as loading control. Quantification was performed using ImageJ. Data were obtained from the same biologic replicate as data shown in panel A and are representative of 3 independent experiments. (D) Protein expression of MCL-1, BCL-2, pERK, and total ERK in HL60 cells treated with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both with or without the addition of the proteasome inhibitor carfilzomib for 4 hours. B-actin levels are given as loading control. Quantification was performed using ImageJ. Data are representative of 3 independent experiments.

Venetoclax-gilteritinib combination reduces ERK and GSK3B phosphorylation and MCL-1 protein levels via proteasomal degradation. (A) Protein expression of MCL-1, BCL-2, pERK, and total ERK in HL60 cells treated for indicated timespan with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. B-actin levels are given as loading control. All western blot images have been cropped for improved clarity and conciseness. Quantification was performed using ImageJ. Data are representative of 3 independent experiments. (B) Protein expression of pGSK3A and B, total GSK3A and B, and MCL-1 in HL60 cells treated for indicated timespan with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. Vinculin levels are given as loading control. Quantification was performed using ImageJ. Data are representative of 3 independent experiments. (C) Protein expression of MCL-1, pMCL-1 S159, and pMCL-1 T163 in HL60 cells treated for indicated time points with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both as analyzed by western blotting. B-actin levels are given as loading control. Quantification was performed using ImageJ. Data were obtained from the same biologic replicate as data shown in panel A and are representative of 3 independent experiments. (D) Protein expression of MCL-1, BCL-2, pERK, and total ERK in HL60 cells treated with 1 μM of gilteritinib, 20 nM of venetoclax, or the combination of both with or without the addition of the proteasome inhibitor carfilzomib for 4 hours. B-actin levels are given as loading control. Quantification was performed using ImageJ. Data are representative of 3 independent experiments.

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