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Fig. 2 | Genome Medicine

Fig. 2

From: A role for the unfolded protein response stress sensor ERN1 in regulating the response to MEK inhibitors in KRAS mutant colon cancers

Fig. 2

Effects of ERN1 inhibition in KRAS mutant human colon cancers. a, b Western blot analysis of ERN1 expression in control cells expressing non-targeting (NT) gRNA and LoVo ERN1KO clones 5B, 6B, and 7B (a) and HCT-116 ERN1KO clones C1, C2, and C3 (b). c, d qPCR analysis of spliced XBP1 mRNA (XBP1s) in control cells expressing non-targeting (NT) gRNA and LoVo ERN1KO clones 5B, 6B, and 7B (c) and HCT-116 ERN1KO clones C1, C2, and C3 (d). Error bars indicate standard deviation calculated from three biological replicates. e Representative colony formation assays of three different ERN1KO clones compared to the non-targeting (NT) gRNA expressing control cells in the KRAS mutant LoVo (top) and HCT-116 colon cancer cells (bottom). Cells were maintained in the indicated range of concentrations of the MEK inhibitor selumetinib (AZD6244) for 10 days, stained and photographed. f, g Live cell proliferation assay (IncuCyte®) of control (NT gRNA) and ERN1KO cells following exposure to the MEK inhibitor AZD6244. Error bars indicate standard deviation of three replicate experiments. h qPCR analysis of spliced XBP1 mRNA (XBP1s) levels following exposure of LoVo cells to increasing concentrations of the ERN1 kinase inhibitor. Error bars indicate standard deviation calculated from three replicate experiments. i Colony formation assay showing the effect of ERN1 kinase inhibitor on the proliferation of KRAS mutant LoVo cells in the presence of the indicated concentrations of the MEK inhibitor AZD6244. j Quantification of spliced XBP1 mRNA (XBP1s) levels following 1 h treatment with 100 nM of ER stress inducer thapsigargin (Tg) in the presence and absence of the ERN1 kinase inhibitor. k Quantification of the mRNA levels of the RIDD target CD59 after 1 h treatment with 100 nM thapsigargin (Tg) in the presence and absence of the ERN1 kinase inhibitor

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