Figure 2

Reversion Assay for putative RNAi factor. (A) (i) Fluorescence imaging of Sf21-gfp reporter cell line (ii) Sf21-gfp reporter cell line transfected with gfp siRNA (iii) Sf21-gfp reporter cell line co-transfected with putative RNAi factor Sf-Dcr-2 specific siRNA with gfp siRNA. (B) Histogram overlay plot shows different gfp expression in transfected Sf21 cell line. Plots depict number of cells (counts) on y axis vs. expression of the gfp reporter (FLH-1) on x axis. Black trace, normal Sf21 cells; green trace, Sf21-gfp reporter cell line; blue trace, siRNA transfected gfp silenced Sf21-gfp cells; red trace, gfp reverted Sf21-gfp cells with putative RNAi factor transfection (a) Sf-Ago-1 (b) Sf-Dcr-2. (C) The bar graph representation of the FACS results for (i) putative core RNAi factors Dicer-2, Argonaute-1, Aubergine, Drosha and (ii) auxiliary RNAi candidates RNA helicase DDX18/HAS1 subfamily, Integrator complex subunit (Int11), Multi Drug Resistance MDR1A transporter and Histone-3 Lysine-4 N-Methyltransferase in Sf21-gfp reporter cell line represented on the x axis, with the percentage of cells expressing gfp on the y axis. Data shown are mean ± SD of three independent experiments.