Additional file 4 of Radiotherapy resistance driven by Asparagine endopeptidase through ATR pathway modulation in breast cancer

Additional file 4. Supplementary Figure 4. Role of AEP in genotoxic tolerance in BC cells. (A) Schematic representation of the systems used to measure the efficiency of DNA repair by different DNA repair mechanisms: classical homologous recombination (DR-GFP), single-strand annealing (SA-GFP) and non-homologous end-joining (EJ5-GFP). (B) qPCR analyses of the mRNA expression levels of AEP and PPP1R10 in control (EV, empty vector) and AEP shRNA (left panel) or control (NT, non-targeting) and AEP siRNA KD (right panel) MDA-MB-231 cells. (C) Co-immunoprecipitation assay of WDR82 using anti-PPP1R10 in control and AEP KD MDA-MB-231 cells. In red box cropped, overexposed WDR82 immunoblot. (D) In vitro digestion of PPP1R10 using recombinant AEP at pH7.2 for 3 hours. (E) Cell death in control and AEP KD MDA-MB-231 after 48 hours irradiation using 0, 2, 5 or 10 Gy. (F) Cell death in non-transfected vs AEP overexpressing HCT116 cells after 24- or 48-hours irradiation with 0 or 5 Gy. * p<0.05, ns: no significant.