Fig. 2: Deletion of ATM using CRISPR/Cas9, but not protein depletion by siRNA knockdown, sensitises PDAC cells to ATR inhibition.

a ATM protein expression in human cell lines at the start (T0) of the drug sensitivity assay (48 h post-transfection) and at the 72-h assay endpoint (120 h post-transfection). Two different “exposures” of the ATM blot are shown (by adjusting brightness of the IRDye image in LiCor Image Studio). Percentage knockdown values vs siCTR are displayed, derived using the LiCor Image Studio quantification software. b AZD6738 dose–response curves of human lines, having been transfected with either a non-targeting siRNA control pool (siCTR) or with an ATM-targeting siRNA pool (siATM). Assay duration was 72 h. Each point represents the mean ± SEM of three independent experiments. c Thirty minutes post 6 Gy of γ-irradiation (IR), MIA PaCa-2 single-cell clones from a CRISPR/Cas9 ATM knockout pool were harvested for immunoblot analysis to determine the ATM status. Clones B4, B7 and C4 were confirmed ATM-null. d AZD6738 dose–response curves of MIA PaCa-2 CRISPR clones. Assay duration was 72 h. Each point represents the mean ± SEM of three independent experiments.