rabbit anti human bcl 2  (Proteintech)

Journal: Cancer Research

Article Title: Targeting BCL2 with Venetoclax Enhances the Efficacy of the KRAS G12D Inhibitor MRTX1133 in Pancreatic Cancer

doi: 10.1158/0008-5472.CAN-23-3574

Figure Lengend Snippet: BCL2 inhibitor venetoclax enhances the efficacy of MRTX1133 in “high”-confluent collagen cultures. A, PDAC cells (2138, 3213, 1245, and PANC1) in “high”-confluent collagen cultures were treated with MRTX1133 (0.5 µmol/L) or the MEK1/2 inhibitor trametinib (0.1 µmol/L) for 8 hours. The effect on BIM and BCL2 was analyzed by Western blotting. Blots are representative of at least three biological replicates. B, PDAC cells in “high”-confluent collagen cultures were cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 8 hours, and the effect on cell death (c-C3, cleaved-PARP) was analyzed. Blots are representative of at least three biological replicates. C, “High”-confluent collagen cultures of PDAC cells were cotreated with venetoclax (V; 2.5 µmol/L) and MRTX1133 (M; 0.5 µmol/L) for 48 hours, and the effect on cell growth was analyzed. Error bars, ± SD; n = 3 to 4. One-way ANOVA, followed by the Tukey multiple comparison test. D, PDAC cells were transfected with control siRNA or siRNA against BIM for 72 hours, plated in collagen, and treated with MRTX1133 (0.5 µmol/L) for 8 hours. The effect on BIM was analyzed using Western blotting. The transfected cells were also cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 8 hours, and the effect on cell death (c-C3) was analyzed. Blots are representative of three biological replicates. E, “High”-confluent collagen cultures of PDAC cells were cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 48 hours with or without the pan-caspase inhibitor zVAD-FMK (zVAD; 10 µmol/L), and the effect on cell growth was analyzed. Error bars, ± SD, n = 3. One-way ANOVA, followed by the Tukey multiple comparison test. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The following antibodies were used at the dilution recommended by the manufacturers: pERK1/2 (#9101, Cell Signaling Technology, RRID:AB_331646), total ERK1/2 (#9102, Cell Signaling Technology, RRID:AB_330744), mouse BIM (#2933, Cell Signaling Technology, RRID:AB_1030947), mouse BCL2 (554218, BD Pharmingen, RRID:AB_395311), cleaved caspase-3 (c-C3; #9664, Cell Signaling Technology, RRID:AB_2070042), human BCL2 (#3498, Cell Signaling Technology, RRID: AB_1903907), PARP (#9542, Cell Signaling Technology, RRID:AB_2160739), HSP90 (#4877, Cell Signaling Technology, RRID:AB_2233307), and GAPDH (MAB374, MilliporeSigma, RRID:AB_2107445).

Techniques: Western Blot, Comparison, Transfection, Control