Fig. 4.

Cx43 HCs reduce intracellular ROS accumulation and increase intracellular GSH level. (A and B) HLE-B3 transfected with Silencer™ Negative Control or Cx43 siRNA were treated with 0.3 mM H₂O₂ for 1 h or 63 mJ/cm² UVB radiation for 30 min. Intracellular ROS and GSH levels were detected by Carboxy-DCFDA (A) or Thioltracker™ (B) fluorescence, respectively. The data are presented as the mean ± SEM. (n = 3). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (Two-way ANOVA). Scale bar: 50 μm. (C and D) HLE-B3 cells were pre-incubated with Cx43E2 antibody for 30 min before H₂O₂ treatment or UVB radiation for various time periods, and followed by incubating with 10 μM Carboxy-H₂DCFDA (C) or Thioltracker™ (D) for 30 min. The data are presented as the mean ± SEM. (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 compared to non-H₂O₂ or UVB treated control (Two-way ANOVA). #, P < 0.05 compared to without Cx43E2 antibody control group (Two-way ANOVA). At least three microphotographs of fluorescence fields were taken under a microscope (Keyence BZ-X710) using a 20X objective and a GFP filter. The average pixel density of 30 random cells was measured using NIH ImageJ software.