ShRNA inhibit gene expression of HBV strains with different genotypes in vitro The levels of cytoplasmic HBV pg/pc RNA (3.5 kb) and HBV DNA in cultured supernatants were

determined by realtime RT-PCR/PCR and presented in Figure 3. The pg/pc RNA level of five HBV strains with different genotypes were reduced by 58%~93% in B245(69%~93%), B376(59%~91%), B1581(67%~90%) and B1789(58%~88%) treatments, while the HBV DNA level observed in supernatants was decreased by 77%~99% in these shRNA plasmid treatments (B245: 83%~99%, B376: 79%~99%, B1581:88%~98%, B1789: 77%~99%). Figure 3 SiRNAs inhibit RNA and DNA expression of HBV strains with different genotypes in Huh7 cells. The histogram show the cytoplasmic HBV pg/pc RNA levels (A, B, C, D, E) and extracellular this website HBV DNA (F, G, H, I, J) of five HBV strains with genotypes Ae(N10), Ba(C4371), C1(Y1021), D1(Y10) and I1(W29) in Sepantronium solubility dmso treated shRNA plasmids, treated pSUPER vector, and non-treated Huh7 cells. In addition, the extracellular and intracellular antigen levels in Huh7 cells that were co-transfected with HBV and shRNA plasmids were also determined (Figure 4). In the shRNA-treated Huh7 cells, the average extracellular HBsAg expression level of all five HBV strains decreased by 1.66 ± 0.36 logs. The average intracellular HBsAg expression level decreased by 1.47 ± 0.33 logs, while the extracellular HBeAg levels decreased by 1.04 ± 0.23 logs, and the intracellular HBcAg levels by 1.71 ±

0.49 logs. The effect of the siRNA treatment on HBeAg levels was weaker than that on the HBsAg or HBcAg levels (P < 0.001, Figure 5). VX 770 Figure 4 SiRNAs inhibit viral antigens expression of HBV strains with different genotypes in Huh7 cells. (A, B, C, D) Extracellular HBsAg, intracellular HBsAg, extracellular HBeAg, and intracellular HBcAg expression levels of HBV N10(Ae), respectively. (E, F, G, H) Extracellular HBsAg, intracellular

HBsAg, extracellular HBeAg, and intracellular HBcAg expression Bay 11-7085 levels of HBV C4371(Ba), respectively. (I, J, K, L) Extracellular HBsAg, intracellular HBsAg, extracellular HBeAg, and intracellular HBcAg expression levels of HBV Y1021(C1), respectively. (M, N, O, P) Extracellular HBsAg, intracellular HBsAg, extracellular HBeAg, and intracellular HBcAg expression levels of HBV Y10(D1), respectively. (Q, R, S, T) Extracellular HBsAg, intracellular HBsAg, extracellular HBeAg and intracellular HBcAg expression levels of HBV W29(I1), respectively. Figure 5 Comparing the RNAi-induced silencing effect on different viral markers. Data were displayed the average antigen level of the 4 siRNAs reduced for five HBV strains. “”Ex”" = Extracellular and “”In”" = Intracellular. The Mann-Whitney test was used to assess the difference. An asterisk represents a statistical difference of P < 0.01 in comparison with the other markers (Ex HBeAg vs. Others P < 0.001, Ex HBsAg vs. In HBsAg P = 0.05, Ex HBsAg vs. In HBcAg P = 0.82, In HBsAg vs. In HBcAg P = 0.10.