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. 2015 Oct 8;10(10):e0139036. doi: 10.1371/journal.pone.0139036

Fig 5. Enhancer compounds facilitate concentration-dependent increase in synthesis of −1-frame HIV-1 protease relative to the 0-frame gag protein.

Fig 5

H9 cells, either infected with HIV-1 IIIB or uninfected as a control, were treated with enhancer compounds A1 and A3 (at 1, 5 and 20 μM) and a DMSO solvent control and cells were harvested at 48 h for analysis of protein expressed in the 0-frame, followed by the appropriate secondary antibody/detection reagent. The filter was then stripped and re-probed with anti-GAPDH for lane-to-lane normalization (A). On a separate gel the protease enzyme (−1-frame) was detected using an anti-protease antibody together with secondary antibody. The filter was stripped and re-probed as before with anti-GAPDH for lane-to-lane normalization (B). The intensity of the developed bands was quantitated (C), and the frameshift efficiency calculated for each compound at each concentration and for the DMSO control. These were normalized to the GAPDH intensities, and compared with the DMSO control.