Fig. 4: BS-DRL1 functions by interacting with HMGB1.

a Interaction of HMGB1 and BS-DRL1 in primary neurons was confirmed by RNA immunoprecipitation (RIP) with HMGB1 antibody following by RT-qPCR, U1 RNA was used as a negative control. Data are presented as mean ± SD. n = 3. b RIP-(RT-qPCR) showed an increased interaction of HMGB1 and BS-DRL1 in neurons upon DNA damage. Primary neurons were treated with DMSO or ETO for 1 h and harvested for following experiments. Data are presented as mean ± SD. n = 3. c, d DNA damage in HMGB1 KD neurons was evaluated by γH2AX immunostaining. Primary neurons infected with indicated shRNAs virus were treated with vehicle or etoposide for 1 h and stained with γH2AX and HMGB1 antibodies. Scale bar: 10 μm. Data are presented as mean ± SD. n = 85 neurons. ****p < 0.0001. AU, arbitrary units. e The level of γH2AX in neurons expressing HMGB1-shRNA was measured by western blot analysis. Primary neurons infected with indicated shRNAs virus were treated with vehicle or ETO for 1 h before harvesting for Western blotting with indicated antibodies. Data are presented as mean ± SD. **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 3 biologically independent experiments. f, g Comet assay to measure the DNA damage accumulation in neurons. Primary neurons infected with indicated shRNA virus were treated with vehicle or ETO for 1 h and then proceeded for comet assay. Scale bar: 50 μm. Data are presented as mean ± SD. n = 63 neurons. ****p < 0.0001, **p < 0.01, ns not significant, AU arbitrary units.