Fig. 2

Stable knockdown of MCL-1 expression reduces ESCC cells proliferation in vitro. a Validation of the MCL-1 knockdown by shRNAs in ESCC cells. KYSE150 and KYSE510 cells were transduced with lentiviral short hairpin RNAs targeting GFP (shGFP) and MCL-1 (shMCL-1#1 and #2). The knockdown of MCL-1 expression was confirmed by Western blot analysis 3 d after shRNAs transduction. β-actin was used as a loading control. b Knockdown of MCL-1 attenuates KYSE150 and KYSE510 anchorage-dependent cell growth. WST-1 assays were performed as described in Materials and Methods. All experiments were performed in triplicate with at least two independent experiments. *p < 0.05, significant difference compared with the shGFP cells at 72 h. c Knockdown of MCL-1 attenuates KYSE150 and KYSE510 anchorage-independent cell growth. Soft agar assays were performed as described in Materials and Methods. **p < 0.01, significant difference in colony formation compared with the shGFP cells