Fig. 2

MCL-1 is required for the MSLC. a The MCL-1 mRNA level in MSLC transfected with MCL-1-siRNA. At 24 h after transfection, the mRNA level of the cells was evaluated using quantitative real-time PCR (**p < 0.01). The MCL-1-siRNA-scrambled was used as a control. b The knockdown of MCL-1 expression at the protein level in MSLC by MCL-1-siRNA. c MCL-1 silencing inhibited the expression of stemness-associated genes in MSLC. At 24 h after transfection of MCL-1-siRNA in MSLC, quantitative real-time PCR was used to evaluate the expression levels of stemness associated genes (**p < 0.01). d MCL-1 knockdown reduced the tumorsphere formation capacity of MSLC. The expression of MCL-1 was silenced by siRNA in MSLC. Seven days after cell seeding, the tumorsphere numbers of MSLC were examined. Control miRNA was used as a control. e The impact of MCL-1 silencing on the MSLC and non-MSLC viability (**p < 0.01). f Knockdown of MCL-1 induced MSLC apoptosis but not in non-MSLC. The MSLC and non-MSLC were treated with MCL-1-siRNA. Twenty-four hours later, apoptosis was examined with the caspase 3/7 activity assay (*p < 0.05). g The detection of apoptosis using annexin V assay. Apoptosis of MSLC and non-MSLC was evaluated by flow cytometry at 24 h after the transfection of MCL-1-siRNA (**p < 0.01)