Fig. 3.

Stable inhibition of mixed-lineage leukemia protein 4 (MLL4) promotes the proliferation in non–small cell lung cancer cells. (A, B) A549 cells and SK-MES-1 cells were transfected with CRISPR activation (CRISPRa)-Scramble or CRISPRa-MLL4 gRNA or the indicated specific shRNAs or/and expression constructs for EdU assays. (C, D) A549 cells and SK-MES-1 cells stably infected with CRISPRa-Scramble or CRISPRa-MLL4 gRNA or the indicated specific shRNAs or/and expression constructs in culture media for 10 days prior to being stained with crystal violet. Representative photos and statistical analyses are shown. (E) The activation and knockdown efficiencies of MLL4 were confirmed by quantitative real-time PCR. The overexpression of PIK3IP1 was confirmed by western blot analysis. Each bar represents the mean±standard deviation for triplicate measurements. *p < 0.05 and **p < 0.01 (two-tailed t test).