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. 2024 May 15;19(6):1243–1249. doi: 10.1021/acschembio.4c00151

Figure 3.

Figure 3

Decrease of NAD RNA capping leads to increased HIV-1 infectivity. (A) Cleavage positions of the NAD RNA cap by the three known decapping enzymes Nudt12, Nudt16 and DXO. (B) In vitro DXO cleavage of NAD-U1 and TMG-U1 at various time points time (0, 10, 30, 60, 120 and 240 min)). (C) Quantification of DXO decapping activity based on PAGE gels (experiments performed in triplicate, error bar represents the standard deviation of the mean). (D) Western blot analysis of DXO and GAPDH (as a control) in control cells and cells with overexpressed (DXO OE). (E) HIV-1 infectivity determined in control cells and cells with overexpressed DXO (DXO OE) (ANOVA test). (F) LC–MS quantification of the NAD RNA cap level in sRNA isolated from control cells and cells with overexpressed DXO (DXO OE) (t-test). (G) LC–MS quantification of the NAD RNA cap level in pulled-down U1 RNA (t-test). All the experiments were performed at least in biological duplicates.