Fig. 2

HMGB1 released from the dying cells in vitro and in vivo. a Release of HMGB1 in the supernatants by 12 Gy-treated Panc-1 and SW1990 was measured by ELISA at the indicated time points (0, 6, 12, 24, and 48 h). b Release of HMGB1 in the supernatants by various radiation doses (0, 4, 8, and 12Gy) treated Panc-1 and SW1990 cells were measured by ELISA. c Western blot analyzing the correlative protein level of Cleaved Caspase-3 and HMGB1 in the supernatants at various time following 12 Gy-treated Panc-1 and SW1990. β-Tubulin was a loading control. d Confocal immunofluorescence analysis demonstrating co-localization of Caspase3 positive cells (green) and HMGB1-positive cells (red) in the xenograft tumors following radiotherapy. Magnification: × 20 and × 60. Experiments were repeated three times and the data were expressed as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001