HMGB1-induced α-SMA expression mediated by RAGE.

<p>Cells were treated with 1, 10, 100 ng/mL HMGB1 for 18h. TLR2, TLR4, and RAGE were detected by immunofluorescence staining (A) and quantification of TLR2, TLR4, and RAGE performed using ImageJ software (B). Cells were transfected with 3 MOI lentiviral based shRNA targeting TLR2, TLR4 and RAGE mRNA for 48h. Cells were collected for mRNA analysis by Real-time Quantitative PCR (C). Transformed cells were added with HMGB1 100ng/ml for further 16 h. Cells were collected and analyzed protein expression by Western Blotting. (D) TLR2 and α-SMA; (E) TLR4 and α-SMA; (F) RAGE and α-SMA. Quantification of TLR2, TLR4, and RAGE (G) and α-SMA (H) performed using Image J software. Data are expressed as mean ± S.E.M. (n ≥ 3) #p<0.01 as compared with NC group. *p<0.05 as compared with mock group (lentiviral vector negative control). a*p<0.05 as compared with NC treated with HMGB1 100 ng/mL. b*p<0.05 as compared with mock group treated with HMGB1 100 ng/mL.