Fig. 5
Co-expression of AGO2 enhances shRNA-mediated knockdown of endogenous genes. a Design of a set of shRNAs targeting human ID1 or P65 mRNA at different regions. The number indicates the initial position of siRNA in the coding region of genes; b a lentivirus harboring U6-driven shID1 and CMV-driven AGO2 or its control was used to infect HeLa cells twice and selected with puromycin for 1 week. Representatives of Western blots using anti-ID1 or anti-ID2 antibodies were shown. β-actin was used as a loading control; c HeLa cell proliferation after silencing of ID1. The same number of stably selected HeLa cells expressing shRNA with or without AGO2 protein were seeded on 96-well plates at a density of 1 × 104 cells per well. After 24 h culture, cell proliferation was determined. The results shown are mean ± SD from four independent experiments. *P < 0.05 vs shCon, ***P < 0.001 vs shCon; d Western blot showing the knockdown of P65 in HeLa cells with a lentivirus-based shRNA with or without AGO2 co-expression. shCon was used as a negative control. β-actin was used as a loading control