Fig. 1

Pamidronate suppresses cell death and reverses the altered expression of caspase 3 and its cleaved in bilirubin-impaired SHED. a A scheme of pamidronate treatment of bilirubin-impaired SHED. SHED were cultured with 0 μM bilirubin (B0), 50 μM bilirubin (B50), and 50 μM bilirubin plus 10 μM pamidronate (B50+PAM) under serum-depleted condition for 3 days. b Cell viability analysis was performed after 3 days of the culture. c, d Cell death assays. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining was performed after 3 days of the culture (c). Flow cytometric assay with Annexin-V (AV) and 7AAD staining was performed after 3 days of the culture (d). e–h Sequential expression of caspase 3 (CAS3) and the cleaved caspase 3 (Cleaved CAS3) was analyzed at the indicated time by western blot analysis. Representative images of western blotting were shown (e). Results were shown as the representative expression of CASP3 to beta-actin (ACTB) (f), Cleaved CAS3 to ACTB (g), and Cleaved CAS3 to CAS3 (h) at each time point in each group. b–d, f–h n = 5 for all groups. Statistical analysis was performed as described in the “Methods” section. Graph bars showed the means ± SEM. b–d *P < 0.05, **P < 0.01, and ***P < 0.005. f–h *P < 0.05, **P < 0.01, and ***P < 0.005 (vs. 0 h in each group). NS, no significance