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Fig. 5 | Retrovirology

Fig. 5

From: The RNA surveillance proteins UPF1, UPF2 and SMG6 affect HIV-1 reactivation at a post-transcriptional level

Fig. 5

SMG6 overexpression leads to attenuated reactivation of HIV-1. a J-Lat 10.6 cells were either mock transfected or transfected with HA-SMG6 and were uninduced (DMSO) or reactivated (PMA). Reactivation, monitored by GFP production, was quantified by flow cytometry and the percentages of reactivation were normalised to the mock-PMA reactivated condition. Error bars represent the standard deviation from three independent experiments with at least 10,000 cells counted per treatment. Asterisks represent statistically significant difference between groups (Two-way ANOVA; p < 0.01). b Cell lysates were run on acrylamide gels and SMG6 and pr55Gag protein levels were detected by Western Blotting. c Example dot plot depicting vRNA expression in mock PMA and SMG6 PMA conditions using FISH-Flow technique. d The % of vRNA expressing cells were quantified. Error bars represent the standard deviation from three independent experiments. Asterisks represent statistically significant difference between groups (One-way ANOVA; p < 0.05). e MFI of the vRNA signal were quantified. Asterisks represent statistically significant difference between groups (student’s t-test; p < 0.05). f J-Lat cells were mock transfected or transfected with HA-SMG6, HA-SMG6-mEBM, HA-SMG6-m14-3-3 or HA-SMG6-mPIN and reactivated with PMA. Cell lysates were run on acrylamide gels and SMG6 and pr55Gag protein levels were detected by SDS-PAGE followed by Western Blotting. g Reactivation in the above conditions was quantified and the percentages of reactivation were normalised to the mock PMA reactivated condition. Error bars represent the standard deviation from three independent experiments with at least 10,000 cells counted per treatment. Asterisks represent statistically significant difference between groups (One-way ANOVA; p < 0.05)

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