Figure 4
From: Mechanisms and consequences of ATMIN repression in hypoxic conditions: roles for p53 and HIF-1
Loss of ATMIN contributes to the hypoxia-mediated repression of DNA repair and a decrease in DYNLL1 expression.
(A) RKO cells transfected with either Scr siRNA or ATMIN siRNA were exposed to 20 h of varying oxygen concentrations (21%, 2% and <0.1% O2) followed by 1 h of MMS treatment (0.1–0.5 mM as indicated) at the same oxygen tension. Cells were then reoxygenated and the media replaced. Colonies were allowed to form at 21% O2 over a period of 8–10 days. Mean surviving fractions are shown ± SD from three independent experiments. (B) RKO cells were transfected with either Scr or ATMIN siRNA and expression of ATMIN and DYNLL1 mRNA was analyzed by qPCR at 48 h post transfection. (C) DYNLL1 mRNA expression was analyzed by qPCR in RKO cells exposed to 16 h of varying oxygen concentrations (21%, 2% and <0.1% O2). (D) RKO cells were transfected with pMSCV-ATMIN vector or an empty vector control. 24 h later cells were transferred to hypoxic conditions (<0.1% O2) for 8 h. qPCR was then carried out for DYNLL1. (E) DYNLL1 mRNA was analyzed by qPCR in RKO cells transfected with either Scr or p53 siRNA and exposed to 16 h of hypoxia (<0.1% O2). All qPCR bar graphs show mean mRNA expression ± SD from three independent experiments. (F,G) Expression of DYNLL1 (Log10 conversion) in the breast (E) and lung (F) TCGA datasets is shown against hypoxia-inducible p53 signature (Log10 conversion). One-tailed p value is shown on each graph for each Pearson r (correlation coefficient).
