Sequencing of Argonaute-bound microRNA/mRNA hybrids reveals regulation of the unfolded protein response by microRNA-320a
Fig 5
miR-320a activates ATF4 during ER stress.
(A) RT-qPCR quantification of spliced(s)-XBP1, BiP, and CHOP mRNA in HCT116 cells transfected with miR-320a mimic. Data are shown as the average of three biological replicates with a standard deviation and individual data points. All three mRNAs were normalized to HPRT. Experiments were performed in technical triplicate. (Student’s t-test): ns = P > 0.05, * = P ≤ 0.05, ** = P ≤ 0.01. (B) Western blots of ATF4 in miR-320a mimic-transfected WT cells with thapsigargin (Tg), tunicamycin (Tm) or histidinol (HisOH) treatment. (C) Western blots of ATF4 in miR-320a antagomir-transfected HCT116 with Tg treatment. Western blots of Atf4 in miR-320a mimic-transfected MEF WT (D) and Perk -/- cells (E) with Tg treatment. In all western blots, GAPDH was used as a loading control. Quantitation of three biological replicates with mean and standard deviation is shown with individual data points. (Student’s t-test): ns = P > 0.05, * = P ≤ 0.05, **** = P ≤ 0.0001.