Figure 1
From: HMGB1 in the pathogenesis of ultraviolet-induced ocular surface inflammation
UV-induced nucleo-cytoplasmic translocation of HMGB1 in human conjunctival epithelial Chang cells. (a) Chang cells were transiently transfected with an HMGB1-GFP plasmid and then irradiated with 20 mJ/cm2 of UV followed by 8-h incubation. The number of cytoplasmic GFP-positive cells was counted among the 100 GFP-positive cells. (b and c) An immunofluorescence assay was performed to observe the UV-induced translocation of endogenous HMGB1. Endogenous HMGB1 was immunostained using an anti-HMGB1 antibody and a secondary anti-rabbit Alexa Fluor 594 antibody. Cytosolic proteins were fractionated from the lysates, and a western blot assay was performed. (d) Chang cells were irradiated with 20 mJ/cm2 of UV and incubated for further 8 h. The HMGB1 level in the culture supernatant was then evaluated by western blot. Relative band intensity was measured using the ImageJ program. Values shown are mean±S.E. *P<0.05 (n=3)
