Figure 5: Effect of HMGB1 Suppression on DNA Damage and DDR After Ang II Stimulation

(A, B) Representative images and analysis of immunostaining of nuclear foci of γ-H2AX (green), α-actinin (red), and DAPI (blue) of NRCMs transfected with small interfering HMGB1 (siHMGB1) or nonspecific control small interfering ribonucleic acid (siRNA) and then treated with saline or Ang II for 24 h. The foci number of γ-H2AX per NRCM was counted. Scale bars = 20 μm. n = 6 in scramble saline group; n = 6 in scramble Ang II group; n = 5 in siHMGB1 saline group; n = 6 in siHMGB1 Ang II group. (C, D) Representative images and analysis of Western blots of γ-H2AX. NRCMs transfected with siHMGB1 or nonspecific control siRNA and then treated with saline or Ang II for 24 h (n = 6 per group). (E, F) Representative images and analysis of immunostaining of nuclear foci of p-ATM (red), α-actinin (green), and DAPI (blue) of NRCMs transfected with siHMGB1 or nonspecific control siRNA and then treated with saline or Ang II for 24 h. The foci number of p-ATM per NRCM was counted. Scale bars = 20 μm. n = 5 in scramble saline group; n = 4 in scramble Ang II group; n = 5 in siHMGB1 saline group; n = 4 in siHMGB1 Ang II group. (G, H) Representative images and analysis of Western blots of p-ATM and total ATM. NRCMs transfected with siHMGB1 or nonspecific control siRNA and then treated with saline or Ang II for 24 h (n = 3 per group). Results are presented as the mean ± SE. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations as in Figures 1, 2, 3, and 4.