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Fig. 4 | Retrovirology

Fig. 4

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

Fig. 4

UPF2 overexpression inhibits the reactivation of HIV-1 in J-Lat cells. J-Lat 10.6 cells were either mock transfected or transfected with Flag-UPF2 and were uninduced (DMSO) or reactivated (PMA). a 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 < 0.05). b Cell lysates were run on SDS-PAGE gels and UPF2 and pr55Gag protein levels were detected by Western Blotting. c Gating strategy to detect UPF2 overexpressing cells by detecting UPF2 mRNA levels by FISH-Flow. d Of the UPF2-mRNA expressing cells gated for in (c), 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 Example dot plot depicting vRNA expression in mock transfected and UPF2 overexpressing populations using FISH-Flow technique. f J-Lat cells were mock transfected or transfected with FLAG-UPF2 or FLAG-UPF2-1-1096. Cell lysates were run on acrylamide gels and UPF2 and pr55Gag protein levels were detected by Western Blotting. g J-Lat cells were mock transfected, transfected with FLAG-UPF2 or co-transfected with FLAG-UPF1 or FLAG-UPF1-LECY. Cell lysates were run on acrylamide gels and UPF2, UPF1 and pr55Gag protein levels were detected by Western Blotting. h Reactivation in the form of GFP expression was quantified in cells transfected as in (f) and (g) 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 < 0.0001)

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