Figure 4

HIV-1 ARM independently confers Dicer dependent RNAi suppressor function. ARM was fused to a reporter protein RFP (ARM RFP). HEK 293 T cells were co-transfected with an expression plasmid for GFP and Dicer dependent shRNA against GFP (0.5 μg) along with RFP vector/ARM RFP. (A) shows a histogram plot (Events on Y axis and expression of GFP (FL1) on X axis) of suppressor assay (as described in 3D) where RSS activity is independently conferred by fusion of ARM to the reporter protein (RFP). The bar graph in (B) shows the quantification of the histogram. (C) shows flow cytometric analysis (Histogram plot showing number of cells (counts) on Y-axis versus the expression of GFP reporter (FL1) on X-axis) and corresponding fluorescence microscopy images of the suppressor assay of ARM-Reporter in the presence and absence of 30 μM Adox (Adenosine, periodate). The bar graph in (D) shows the quantification of the histogram. (E) show the ARM mediated reversal of Dicer-dependent-silencing assay based on luciferase reporter with a significant p value (< 0.001).