Fig. 5

The ATM inhibitor decreased the expression of CLCA2 and inhibited cell apoptosis induced by TP63-mut protein expression (A)293FT cells were transfected with empty vector (NC), TP63-WT, TP63-mut, pGL3-Basic-CLCA2 prom, and pRL-TK plasmids, and treated with the ATM inhibitor (ATMi). Luciferase assay results demonstrated that the expression of the CLCA2 promoter was significantly reduced in TP63-mut + ATMi cells compared to that of TP63-mut cells, indicating that the ATM inhibitor reduced CLCA2 expression to regulate the effect of TP63-mut protein. (B) 293FT cells were transfected with empty vector (NC), TP63-WT, TP63-mut, pGL3-Basic-CLCA2 prom, and pRL-TK plasmids, and treated with ATMi and siCLCA2. The RT-qPCR data further confirmed that ATMi inhibited CLCA2 expression, with a more pronounced effect observed in cells treated with both ATMi and siCLCA2. (C-D) The combination of ATMi and siCLCA2 inhibited the pro-apoptotic effect of the TP63-truncating mutation. Annexin-V was analyzed using Flowjo_v10.8.1 and GraphPad Prism