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. 2015 Jun 1;65(7):1151–1164. doi: 10.1136/gutjnl-2014-308270

Figure 2.

Figure 2

Figure 2

B-cell CLL/lymphoma 3 (BCL-3) expression inhibits apoptosis in colorectal cancer cells. (A) Suppression of BCL-3 expression significantly decreased the total cell yield and increased cell death compared with negative small interfering (si)RNA-transfected HCT116 cells 72 h post transfection. Suppression of BCL-3 protein expression was confirmed by western analysis. (B) BCL-3 overexpression significantly increased the total cell yield and decreased cell death compared with control plasmid-transfected SW480 cells 72 h post transfection. Overexpression of BCL-3 was confirmed by western analysis. (C) Treatment with 5 μM QVD (pan-caspase inhibitor) for 72 h reduced cell death significantly in control and BCL-3 knockdown HCT116 cells confirming induction of apoptosis following suppression of BCL-3 expression (***p<0.001, **p<0.01, NS, non-significant). Western analysis confirmed suppression of BCL-3 expression in QVD-treated HCT116 cells. Fluorescence-activated cell sorting analysis showed a significant increase in apoptosis (determined as sub-G1 peak, arrows) from 11.4% to 27.5% following BCL-3 suppression. In cells treated with QVD, apoptosis is inhibited in both control and BCL-3 knockdown cells (arrows). Results are representative of a single experiment performed in duplicate, assessed 72 h post transfection. (D) The decrease in cell death on stable overexpression of wildtype (wt)BCL-3 was not detected when the ANK M123 mutant (mut)BCL-3 was stably overexpressed compared with control plasmid-transfected SW480 cells 72 h after seeding. Overexpression of BCL-3 and mutBCL-3 was confirmed by western analysis.