TNRC6A is essential for ras-induced DDR and p53 activation.
(A–C) BJ cells transduced with miR-30 or vector (Ctrl) or shRNA for TNRC6A or Dicer or a scrambled sequence (shSC) and HaRasV12 (Ras) or vector (WH) were stained for 53BP1- or γ-H2AX-foci and photographed (A). % of cells with > 5 53BP1- (B) or γ-H2AX (C)-foci (arrows) were quantified.
(D–E) Western blot of BJ cells transduced with miR-30c-1 or vector (Ctrl) (D) or shRNA for TNRC6A or a scrambled sequence (shSC) (E) and then with HaRasV12 (Ras) or vector (WH) on day 8 post Ras-transduction.
(F) BJ cells with a stable p53-dependent luciferase reporter (PG-Luc) were transduced with miR-30 or vector (Ctrl) (left) or shRNA for TNRC6A or Dicer or a scrambled shRNA (shSC) (right) and then with HaRasV12 (Ras) or vector (WH). Luciferase activity was determined 7 days later and normalized to protein concentrations.
(B–C, F) Values are mean±SD for triplicates. * p < 0.05 vs Ctrl or shSC by Student t test.