The protective effect from rI/R by E15 in the timeline. Necrosis was determined by high-mobility group box-1 (HMGB1) immunostaining assay. (A) Sham group at 48 h; (B) rI/R-veh at 6, 24, and 48 h after rI/R (upper panel) and E15-treated groups at the same times; (bottom panel). (C) Box-and-whisker plot (median, first-third quartile, minimum- maximum value) with the percentage of positive cells. Representative necrotic cells are shown with red arrows and their nuclei stained with DAPI (blue). * p < 0.05 compared to rI/R-veh α
p < 0.01 compared to rI/R-veh and rI/R-E15 groups, at 48 h. Scale bar = 100 μm.