Fig. 6

Effects of silencing ATG4B on mobility and chemosensitivity in human colorectal cancer cells. (A) HCT116 cells were transfected with 10 nM scramble siRNA (siCtrl) or three individual siRNA against ATG4B (siATG4B #1, #2 and #3) or pooled siRNA (#P) against ATG4B for 72 h and examine knockdown efficiency by immunoblotting. (B) The ATG4B silenced cells were observed in left panel and measured for cell viability by CellTiter Glo. (C) ATG4B silenced CRC cells were cultured in migration insert and (D) Transwell chamber to quantify the migratory distance and invaded cells, respectively. (E) CRC cells were silenced with 10 nM scramble or siRNA against ATG4B for 48 h and treated with chemotherapeutic drug irinotecan (CPT, 1 µM) or oxaliplatin (OXI, 10 µM) for 24 or 48 h. The cell viability was measured with CellTiter Glo. (F) The siRNA transfected CRC cells were cultured as spheres and treated with chemotherapeutic drug (CPT, 1 µM, OXI, 10 µM) for 2 days as images. The tumor spheres were quantified compared to control sphere. Scale bar: 400 μm. (G) The sphere viability was monitored by LIVE/DEAD staining dyes. The fluorescence was read and quantified. * p < 0.05, ** p < 0.01, *** p < 0.001