Fig. 1

Effects of AGO1, AGO2, DICER or XPO5 overexpression on gene silencing. a Comparison of the silencing activities of two shRNAs against EGFP. HEK293 cells were co-transfected with pENTR/CMV-EGFP, pDsRed2-c1 and U6 promoter-driven shEGFP450, shEGFP417 or an unrelated shCon. Two days after transfection, the cells were then collected for flow cytometry analysis to evaluate EGFP knockdown efficiency by determining green to red (G/R) fluorescence intensity ratio; b, c effects of AGO1, AGO2, Dicer or XPO5 overexpression on the silencing of EGFP with shEGFP417 or shEGFP450. HEK293 cells were co-transfected with shRNA plasmid (shEGFP450, shEGFP417 or shCon), pENTR/CMV-EGFP, pDsRed2-c1 and plasmid expressing AGO1, AGO2, Dicer, XPO5 or empty control vector. Two days after transfection, G/R fluorescence intensity ratio was analyzed by flow cytometry and normalized to the shCon/control group; d representative images from the cells 2 days after transfection of pENTR/CMV-EGFP, pDsRed2-c1 and shGFP450 plasmids in the presence of AGO1, AGO2, Dicer, XPO5 or empty control vector; e G/R fluorescence intensity ratio measured from the cells transfected with shRNAmiR30a-based shEGFP417 (shEGFP417-miR30a) and the plasmid expressing AGO1, AGO2, Dicer, XPO5 or empty vector control. The results shown are mean ± standard deviation (SD) from three independent experiments. *P < 0. 05, ***P < 0.001