Figure 2.
PLX51107 and PLX2853 BETi upregulate the BH3-only protein BIM in Eμ-myc lymphomas. (A) Eμ-myc lymphoma cell lines (AH29, AF47, and Eμ# 4) were treated with qVD (25 μM) for 1 hour to inhibit cell death, followed by PLX5, PLX2, or a DMSO vehicle control (Con) for 24 hours. BIM mRNA expression was then assessed, in triplicate, by quantitative polymerase chain reaction (qPCR) normalized to GAPDH. (B) Eμ-myc lymphoma cell lines (n = 3) were treated as in panel A, and BIM protein expression was assessed after 48 hours. (B) Representative example, with densitometry data normalized to a DMSO control (Con) for 1000 nM PLX5 or PLX2 from each cell line depicted in panel C. (D-E) BCL-2 OE Eμ-myc lymphoma cell lines (n = 3) were treated with 1000 nM PLX5 or PLX2 or a DMSO vehicle control (Con), and BCL-2 protein was immunoprecipitated after 48 hours. Levels of BIM coimmunoprecipitation were then assessed. (D) Representative example of input whole cell lysates (i) and BIM:BCL-2 coimmunoprecipitation (ii) with coimmunoprecipitation densitometry data normalized to baseline demonstrated in panel E. Error bars represent SEM. Statistical analysis was performed using 2-way (A) or 1-way (C-E) ANOVA, and P values were adjusted using the Sidak method or Dunnett’s multiple comparisons tests, respectively. **P < .005, *P < .05.

PLX51107 and PLX2853 BETi upregulate the BH3-only protein BIM in Eμ-myc lymphomas. (A) Eμ-myc lymphoma cell lines (AH29, AF47, and Eμ# 4) were treated with qVD (25 μM) for 1 hour to inhibit cell death, followed by PLX5, PLX2, or a DMSO vehicle control (Con) for 24 hours. BIM mRNA expression was then assessed, in triplicate, by quantitative polymerase chain reaction (qPCR) normalized to GAPDH. (B) Eμ-myc lymphoma cell lines (n = 3) were treated as in panel A, and BIM protein expression was assessed after 48 hours. (B) Representative example, with densitometry data normalized to a DMSO control (Con) for 1000 nM PLX5 or PLX2 from each cell line depicted in panel C. (D-E) BCL-2 OE Eμ-myc lymphoma cell lines (n = 3) were treated with 1000 nM PLX5 or PLX2 or a DMSO vehicle control (Con), and BCL-2 protein was immunoprecipitated after 48 hours. Levels of BIM coimmunoprecipitation were then assessed. (D) Representative example of input whole cell lysates (i) and BIM:BCL-2 coimmunoprecipitation (ii) with coimmunoprecipitation densitometry data normalized to baseline demonstrated in panel E. Error bars represent SEM. Statistical analysis was performed using 2-way (A) or 1-way (C-E) ANOVA, and P values were adjusted using the Sidak method or Dunnett’s multiple comparisons tests, respectively. **P < .005, *P < .05.

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