MTMR 2/3/4/7 regulate the oncogenic KRAS signal output. (A) RNAi was induced by feeding let-60 (n1046) L1 worms through the adult stage with E. coli strain HT115, producing dsRNA to target genes. Presence of the multivulva phenotype was scored using DIC/Nomarski microscopy. 100–200 worms were assayed per RNAi KD. (B–D) T47D cells stably expressing (B) GFP-KRASG12V or (C) GFP-HRASG12V, or (D) WT BxPC3, MIA PaCa-2, and PANC-1 cells were infected with lentiviruses expressing SCR or shRNA targeting MTMR 2/3/4/7, followed by 1 μg/ml puromycin selection. Cells were seeded on a 96-well plate and cultured for 4 days. Complete growth medium was replaced every 24 h, and cell numbers were counted to measure cell proliferation. The graphs show the mean cell proliferation ± SEM relative to that for the control cells (SCR-expressing) from three independent experiments. (E) Cell lysates prepared from D were immunoblotted for ppERK, pAkt (S473), c-MYC, and endogenous KRAS. Values indicate the mean ppERK, pAkt, c-MYC, or endogenous KRAS ± SEM relative to the control cells (SCR-expressing) from three independent experiments. Total ERK, Akt, and actin blots were used as loading controls. Representative blots are shown. Significant differences between control (SCR-expressing) and MTMR KD cells were assessed using one-way ANOVA tests (*P < 0.05, **P < 0.01, ***P < 0.001, #—not significant). SCR, scrambled shRNA. Source data are available for this figure: SourceData F7.