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Figure 2: Gene silencing effects of short interference RNAs against non-structural 5B in stable human hepatoma-7 cell clones. The silencing effects of short interference RNA targeting non-structural 5B gene were analysed by RT-polymerase chain reaction, Western blotting, and real-time polymerase chain reaction. (a and b): mRNA expression of non-structural 5B gene after 24 and 48 h post-transfection with 50nM short interference RNAs compared to mock (polymerase chain reaction 3.1/FlagTAG), positive control (polymerase chain reaction 3.1/FlagTAG/non-structural 5B) and scrambled short interference RNA-treated cells expression. All experiments were performed in triplicate. mRNA expression of GAPDH was used as an internal control both in short interference RNA-treated and untreated cells. L: 100 bp DNA ladder, M: human hepatoma-7 cells with mock transfection, C: Positive control cells. (c) Quantitative real-time polymerase chain reaction showed a relative decrease in mRNA expression of non-structural 5B after 24 and 48 h post-transfection with short interference RNAs as compared to controlled expression (i.e., Mock transfection). Each sample was tested in triplicates. The error bars indicate mean (n = 3) ± standard deviation whereas P* and P^ designated significant differences between connected groups. (d): short interference RNA gene silencing effect at the protein level as determined by Western blotting. M: human hepatoma-7 cells with mock transfection, C: Positive control cells

Figure 2: Gene silencing effects of short interference RNAs against non-structural 5B in stable human hepatoma-7 cell clones. The silencing effects of short interference RNA targeting non-structural 5B gene were analysed by RT-polymerase chain reaction, Western blotting, and real-time polymerase chain reaction. (a and b): mRNA expression of non-structural 5B gene after 24 and 48 h post-transfection with 50nM short interference RNAs compared to mock (polymerase chain reaction 3.1/FlagTAG), positive control (polymerase chain reaction 3.1/FlagTAG/non-structural 5B) and scrambled short interference RNA-treated cells expression. All experiments were performed in triplicate. mRNA expression of GAPDH was used as an internal control both in short interference RNA-treated and untreated cells. L: 100 bp DNA ladder, M: human hepatoma-7 cells with mock transfection, C: Positive control cells. (c) Quantitative real-time polymerase chain reaction showed a relative decrease in mRNA expression of non-structural 5B after 24 and 48 h post-transfection with short interference RNAs as compared to controlled expression (i.e., Mock transfection). Each sample was tested in triplicates. The error bars indicate mean (<i>n</i> = 3) ± standard deviation whereas <i>P</i>* and <i>P</i><sup>^</sup> designated significant differences between connected groups. (d): short interference RNA gene silencing effect at the protein level as determined by Western blotting. M: human hepatoma-7 cells with mock transfection, C: Positive control cells