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Fig. 4 | Virology Journal

Fig. 4

From: Cytosolic sulfotransferase 1A1 regulates HIV-1 minus-strand DNA elongation in primary human monocyte-derived macrophages

Fig. 4

SULT1A1 regulates HIV-1 reverse transcription. a MDMs were treated with control siRNA or SULT1A1-specific siRNA and subesequently challenged with VSV-G pseudotyped NL43-Luc HIV-1 vector. DNA was isolated 24 h post-infection and qPCR was performed using primers that detect early RT DNA products or late RT DNA products compared to the cellular PBGD gene as an endogenous control. The levels of early and late RT products were normalized to the ASN siRNA control. Results shown are from MDMs derived from 6 donors tested twice. b MDMs were transfected with SULT1A1 siRNA 2 or ASN siRNA and total cellular RNA was isolated 24 h post infection with VSV-G pseudotyped NL43-Luc HIV-1 vector. qPCR using primers specific for HIV-1 multiply spliced mRNA (MS RNA, forward and reverse primers span the first and second exons of Tat/Rev, respectively) was performed, and relative MS RNA (normalized to GAPDH) was then normalized to ASN siRNA control. Results shown are from MDMs derived from 6 donors tested twice. c Representative immunoblot showing HIV-1 Vpu and Vif protein levels compared to endogenous Ku86 or GAPDH loading control, respectively, from protein lysate collected 48 h post infection with VSV-G pseudotyped NL43-Luc HIV-1 vector pre-treated with either ASN siRNA control or SULT1A1 siRNA 2. d Quantitative immunoblot analysis using Image Studio software of Vpu and Vif as shown in Fig. 4c. Mean and SD shown, *** p < 0.0005 ** p < 0.005 * p < 0.05 one sample t test. Results shown are from 6 donors assayed twice. Samples with <60 % SULT1A1 knockdown and/or <65 % cell viability were not used in the analysis. All values were compared to ASN control

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