Inhibition of high-mobility group box 1 expression by siRNA in rat hepatic stellate cells

Figure 4 Screening the most effective high-mobility group box 1 siRNA sequence. Total RNA and protein were obtained from hepatic stellate cell-T6 transfected with negative control (NC), mock and three different high-mobility group box 1 (HMGB1) siRNA molecules (shRNAH1, shRNAH2 and shRNAH3). A: Real-time polymerase chain reaction (RT-PCR) for the effect of three different HMGB1 siRNA molecules on HMGB1 mRNA level 48 h after transfection. The expression was normalized against β-actin; B: Semiquantitative analysis of the RT-PCR result; C: Western blotting analyzed HMGB1 protein expression 48 h after transfection; D: Semiquantitative analysis of the western blotting results. Data represent results from one of three similar experiments. Results show that all three HMGB1 shRNA constructs were effective, but shRNAH3 was more efficient in reducing the HMGB1 mRNA and protein levels than shRNAH2 and shRNAH1.