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. 2009 Jul 9;3:63. doi: 10.3389/neuro.17.001.2009

Figure 2.

Figure 2

Effect of different promoters and Ago2 co-expression in RNAi efficacy. HEK293 cells, transfected with pSi and pRe constructs were imaged after 48 or 72 h to evaluate GFP knockdown efficiency by the green to red (G/R) fluorescence intensity ratio. (A) Diagram of the pSi constructs (left) containing U6, enhancer added U6 (EnU6) or H1 promoter driven shGFPs, and the pRe construct (right) for simultaneous GFP and mCherry expression. (B) Representative images from cells 2 days after transfection of pSi constructs containing EnU6 driven shGFPs with or without Ago protein co-expression. (C) G/R fluorescence intensity ratio measured from cells transfected with pSi constructs with or without Ago protein co-expression. The ratio in shGFP-transfected cells was normalized to the averaged G/R ratio from controls with no shRNA expression. shGFPs driven by U6, EnU6 and H1 promoters have comparable degrees of knockdown. However, EnU6-shGFPs with Ago2 co-expression show significant increases in GFP knockdown compared to EnU6-shGFPs alone or EnU6-shGFPs with Ago1 co-expression. Asterisks represent statistical significance (p < 0.05, ANOVA-Scheffe's test).