Skip to main content
. 2020 Jan 27;11(1):68. doi: 10.1038/s41419-020-2263-0

Fig. 5. BCL-G is dispensable for apoptosis induced by IFN-γ and TNF-α in human IEC.

Fig. 5

a–c HT-29 cells transfected with a non-targeting siRNA (siCtrl) or siRNA targeting BCL-G (siBCL-G) were stimulated 48 h later with IFN-γ+TNF-α (1 or 10 ng/ml each) for various time points. a Relative mRNA expression of BCL-GS/L as measured by RT-qPCR at 8 h after cytokine treatment. b Western blots showing levels of BCL-GL and caspase-3 (total and cleaved) at 24 h after cytokine treatment (left). Caspase-3/7 activity as measured at 24 h (right). c Relative cell viability and crystal violet staining of HT-29 cells treated as indicated for 48 and 72 h, respectively. d Western blots showing levels of BCL-GL and caspase-3 (total and cleaved) in wild-type or BCL-G knockout (BCL-GKO) HT-29 cells treated with IFN-γ+TNF-α (10 ng/ml each) for 24 h (left). Caspase-3/7 activity was measured at 24 h (right). e Relative cell viability and crystal violet staining of wild-type or BCL-GKO HT-29 cells treated with IFN-γ+TNF-α (10 ng/ml each) for 72 h. f, g DLD-1 cells were transfected with a tGFP-tagged empty vector (p.Empty) or BCL-GS (p.BCL-GS) or BCL-GL (p.BCL-GL) expression plasmids for the indicated time points. f Relative mRNA expression of BCL-GS/L as measured by RT-qPCR (left) and western blots showing protein levels of BCL-GS/L (right) at 24 and 48 h. g Relative viability of DLD-1 cells at 24 and 48 h after transfection (left) and crystal violet staining of DLD-1 cells transfected for 24 h, followed by treatment with IFN-γ+TNF-α (1 or 10 ng/ml each) for 72 h (right). h Relative viability of HT-29 cells plasmid transfected for 48 h, followed by treatment with IFN-γ+TNF-α (1 or 10 ng/ml each) for 72 h (top). Western blot showing protein levels of BCL-GS/L at 48 h after plasmid transfection (bottom). Data shown are the mean ± S.E.M. of at least n = 3 independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 (two-way ANOVA followed by Tukey’s multiple comparisons test as indicated). For crystal violet staining, grey dotted lines indicate splicing of images. NT — non-treated, wt — wild-type.