310948.fig.004
Figure 4: Efficacy screening of the three designed shRNA via images analysis. pDsRed1-C1-PPARγ vector was transfected as a control ((a1), (b1), and (c1)). The three tested shRNA (sh500, sh614, and sh1006) as pENTR/CMV-GFP/U6-shRNA construct were cotransfected with pDsRed1-C1-PPARγvector. The transduction efficiency was estimated by the level of green fluorescent protein (GFP) expression ((a3), (b3) and (c3)). Shown are representative images of the PPARγ expression (in red) after a 48 h cotransfection. (a1), (b1), and (c1) show high transfection and expression of PPARγ construct vector. (a2), (b2), and (c2) show reduction of PPARγ expression after addition of shRNA construct, while (a3), (b3), and (c3) show efficacy of shRNA transfection as shown by the green color (i.e., GFP). Images were obtained by a fluorescence microscope (Leica, DMI4000B, Germany) at 100x magnification. The images clearly show that the sh1006 had the highest effect on PPARγ vector expression (c2).