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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: J Hepatol. 2018 Mar 9;68(6):1228–1238. doi: 10.1016/j.jhep.2018.01.026

Fig. 2. FGFR inhibition depletes MCL1 expression in a dose- and time-dependent manner leading to cell death.

Fig. 2

(A) After KMCH and KMBC cells were treated with increasing concentrations of LY2874455 (LY) for 24 h, MCL1 mRNA and mitochondrial protein levels were assessed by qPCR and western blotting, respectively. HSP60 was used as a loading control for mitochondrial lysate. (B) KMCH and KMBC cells were treated with LY (5μM) for 4, 8, and 24 hours and MCL1 mRNA and mitochondrial protein levels were examined. Bar graphs represent mean ± SEM for at least three independent experiments. ***p < 0.001 compared to vehicle treatment. (C) KMCH cells were stably transfected with Cas9 and doxycycline-inducible sgRNA targeting MCL1 gene (isgMCL1). Cells were treated with doxycycline (5 μg/mL) for several time points and cell death rate was evaluated by CellTiter-Glo luminescent assay. ***p < 0.001, **p < 0.01. Statistical significance was determined using the two-tailed t test or one-way ANOVA followed by Dunnett’s test.