Fig. 4: DSBs induce SPINDOC expression at both the mRNA and protein levels.

a HEK293T cells were treated with 10 Gy IR and 40 μM Etoposide for one hour before media replacement, and samples were taken at the indicated timepoints for Western blot analysis. Nontreated (NT) cells serve as a control, and DMSO treatment (D) as a solvent control. The results were shown as representative blots from three times independent experiments. b HEK293T cells were treated with DMSO or 10μM CHX and then immediately treated with 10-Gy IR (left panels) or 40μM Etoposide for an hour before replacing media (right panels) and incubated for either 3- or 6 h. Western blot analysis was then performed using the indicated antibodies. The results were repeated three times independent experiments, shown as representative blots. c HEK293T and Hela cells were treated with 10-Gy IR and cultured further for 0.5, 3, and 6 h. Total RNAs were extracted for RT-qPCR analysis of SPINDOC mRNA levels. Graphs represent mean ± SD, n = 3 biologically independent samples. Statistical analysis was performed using one-tailed Student’s t-test with HEK293T IR treatment compared with NT, 0.5 h, P = 0.0320, 3 h, P = 0.0126, 6 h, P = 0.0027, ***P < 0.001.