6 DNA damage-induced p53 downregulates expression of RAG1 | 163 Supplemental Figure 2. (A) Western Blot staining of RAG1, cleaved caspase-3 and KU70, used as a housekeeper, in BV173 cells, treated first with STI571 for 24 hours to induce RAG1/2 expression and Igk recombination. Then, the cells split into 4 groups, a control group, a group treated with p53 stabilizer Nutlin-3 (2.5mM) for 4 hours, a group treated simultaneously with Nutlin-3 (2.5mM) and the pan-caspase inhibitor Z-VADFMK (50mM) for 4 hours and a group treated with caspase-activator Staurosporine (200nM) for 4 hours. (B) FACS staining for propidium iodide showing the % of sub-G1 cells, defined as dead cells, in mouse v-Abl WT cells (A70) treated initially for 72 hours with STI571 to induce RAG1/2 expression and Igk recombination. After 72 hours, the cells were either subjected to 5Gy g-irradiation and left to recover for 2 hours or pre-treated with ATM inhibitor KU55933 for 2 hours prior to the g-irradiation or treated for 4 hours with KU55933 with no irradiation. The DMSO-treated group was taken as a control.
RkJQdWJsaXNoZXIy MTk4NDMw