Figure 2
From: HMGB1 in the pathogenesis of ultraviolet-induced ocular surface inflammation
UV-induced ROS generation resulting in the translocation of HMGB1. (a) Chang cells were treated with 20 mJ/cm2 of UV with or without pretreatment with NAC and Mito-Tempo (Enzo Life Sciences). Approximately, 5 μmol/l of DC-FDA was used to detect ROS generation. Mean fluorescence intensity was measured using FV1000 confocal microscopy. *P<0.05 and **P<0.01 (n=3). (b) Chang cells were treated with 20 mJ/cm2 of UV with or without pretreatment with NAC and Mito-Tempo (Enzo Life Sciences). At 8 h after UV treatment, 8-OHdG levels were measured by ELISA. **P<0.01 (n=3). (c) Chang cells were transfected with the HMGB1-GFP plasmid and incubated for 24 h. UV treatment was performed with or without pretreatment with NAC (10 mM) and Mito-Tempo (Enzo Life Sciences, 50 nM) 1 h before UV radiation. After 8 h, the GFP-tagged HMGB1 signal was visualized by confocal microscopy. The number of cytoplasmic HMGB1-GFP cells was then counted. Values shown are the mean±S.E. *P<0.05 (n=3)
