Figure 5. Metformin inhibited the relocation of HMGB1 in rabbit AF cells induced by LPS. (A–D) H33342 staining. (E–H) anti-HMGB1 antibody staining. (I–L) merged images of the H33342 stained images (A–D) and anti-HMGB1 antibody stained images (E–H). (M–P) the enlarged images of the box areas of the images of (E–H, Q) semi-quantification of positive stained HMGB1 in the nuclei of the cells. More than 95% of the nuclei of the normal cells were positively stained with HMGB1 (I, M, Q). LPS induced HMGB 1 releasing from the nucleus of rabbit AF cells to the cytoplasma (J, N, Q). Metformin blocked the relocation of HMGB1 as evidenced by the concentration of HMGB1 in the nuclei (K, O, Q). *p<0.05 compared to the control group. #p<0.05 compared to the cells treated with LPS. White bars = 100 μm, yellow bars = 25 μm.