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. 2015 Oct 16;35(22):3866–3879. doi: 10.1128/MCB.00528-15

FIG 1.

FIG 1

KSR1 regulates anchorage-independent growth and tumor maintenance in colon cancer cell lines. (A) Immunoblot of KSR1 protein expression in multiple colon tumor cell lines compared to expression in HCECs. (B) qPCR analysis of mRNA levels of KSR1 in colon cancer cell lines relative to levels in HCECs normalized relative to glyceraldehyde-3-phosphate dehydrogenase. (C) Validation of individual siRNA duplexes targeting KSR1. (D and E) Immunoblot of KSR1 and analysis of apoptosis following siRNA-mediated depletion of KSR1 in HCECs and HCT116 cells. (F) Immunoblots of HCT116 cells following shRNA-mediated depletion of KSR1. (G) Quantification of anchorage-independent HCT116 cell growth in soft agar following KSR1 RNAi. Representative bright-field pictures are shown (bottom panel). (H) Immunoblots of KSR1 protein expression (top) and quantification of anchorage-independent HCT116 cell growth in soft agar (bottom) following stable knockdown of KSR1 with a rescued phenotype shown after hairpin-resistant mouse Flag-tagged KSR1 was reintroduced. (I) Xenograft tumor growth of HCT116 cells. ***, P < 0.001; ****, P < 0.0001.