Fig. 6
STAT1 mediates LPS+CS-induced RAW 264.7 cell death by inhibiting the translocation of HMGB1. RAW 264.7 cells were transfected with 100 nM of control siRNA or STAT1-specific siRNA for 48 h and then subjected to 1 μg/ml LPS for 24 h plus 24% elongation stretch for 12 h at a frequency of 30 cycles/min. AZD1480 (5 μM) was given 30 min before LPS stimulation when necessary. a Western blot analysis of STAT1 knockdown in RAW 264.7 cells. b Western blot analysis of the levels of LC3-I, LC3-II in RAW 264.7 cells treated with LPS+CS or/and HMGB1 siRNA or scramble (Ctr) siRNA. c Western blot analysis of the levels of cleaved caspase 3 and total caspase 3 in RAW 264.7 cells treated with LPS+CS or/and STAT1 siRNA or scramble (Ctr) siRNA. d Measurement of the released LDH in RAW 264.7 cells treated with LPS+CS or/and STAT1 siRNA or scramble (Ctr) siRNA. e Western blot analysis of the levels of cytoplasmic HMGB1 in RAW 264.7 cells treated as indicated. β-Actin was loading control for cytoplasmic proteins. f Western blot analysis of total nuclear HMGB1 in RAW 264.7 cells treated as indicated. Lamin B1 was loading control for nuclear proteins. g Western blot analysis of cytoplasmic HMGB1 in RAW 264.7 cells treated as indicated. β-Actin was loading control for cytoplasmic proteins. h Western blot analysis of nuclear HMGB1 in RAW 264.7 cells treated as indicated. Lamin B1 was loading control for nuclear proteins. Data represented the mean ± SEM from at least two independent experiments. *p < 0.05 and **p < 0.01, ***p < 0.001 compared with control or Ctr siRNA + LPS+CS. &p < 0.05 and &&p < 0.01, compared with the LPS+CS
