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. 2018 Mar 22;9(4):436. doi: 10.1038/s41419-018-0402-7

Fig. 5. NRBP1 regulates caspase-dependent intrinsic apoptosis in CRC cells.

Fig. 5

a SW480 and HCT116 cells were treated with Z-VAD-FMK for 1 h, a pan-caspase inhibitor. Twenty-four hours later, apoptotic ratios were analysed by double staining the cells with Annexin V and PI using flow cytometry. Representative images from triplicate experiments are shown (top). Q3-1 necrosis cells. Q3-2 late apoptosis cells. Q3-3 normal cells. Q3-4 early apoptosis cells. Graphs (bottom) show quantitative analysis of total apoptosis. b Western blot was performed to investigate the expression levels of caspase-related proteins (caspase-9, caspase-8, caspase-3) in SW480 and HCT116 cells transduced with lenti-NRBP1 (lenti-GFP was used as a negative control). GAPDH was used as loading control. c Western blot was performed to investigate the expression levels of caspase-related proteins (caspase-9, caspase-8, caspase-3) in RKO and DLD1 cells transduced with shNRBP1. GAPDH was used as loading control. d Bax, Bcl-2, cytosolic cytochrome c and GAPDH (loading control) protein levels were detected by western blot in SW480 and HCT116 cells transduced with lenti-NRBP1 (lenti-GFP was used as a negative control). e Bax, Bcl-2, cytosolic cytochrome c and GAPDH (loading control) protein levels were detected by western blot in RKO and DLD1 cells transduced with shNRBP1. f The effects of Z-VAD-FMK on caspase-related protein in SW480 and HCT116 cells transduced with lenti-NRBP1 or lenti-GFPS were evaluated by western blot analysis (*P < 0.05, **P < 0.01)