ER stress-induced inhibition of cap-dependent translation and loss of MCL1 is PERK-dependent.

<p><b>(A)</b> HCT116 cells were pre-treated for 1 h with the indicated concentration of GSK2606414 before addition of 100 nM Tg for 6 h. Whole cell lysates were separated by SDS-PAGE and analysed by immunoblotting using the indicated antibodies. <b>(B)</b> HCT116 cells were transfected with a dual luciferase reporter construct for assay of CAP/IRES-dependent translation. 24 h post-transfection, cells were pre-treated for 1 h with 100 nM GSK2606414 (GSK) before addition of 2 μg ml^-1 Tm or 1 μM AZD8055 for 24 h. Results shown are the mean ± S.D. luciferase activity within the whole cell lysates of one experiment performed in technical triplicate and are representative of three independent experiments. Statistics shown are the results of Student’s unpaired <i>t</i>-tests; N.S., not significant; *, p < 0.05. <b>(C)</b> HCT116 cells were pre-treated for 1 h with 100 nM GSK2606414 prior to the addition of the indicated concentration of Tm for 24 h. Whole cell lysates were fractionated by SDS-PAGE and analysed by immunoblotting using the indicated antibodies. Results in (A) and (C) are representative of 3 independent experiments.