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. 2022 May 9;12:7570. doi: 10.1038/s41598-022-11230-8

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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APOBEC3A is involved in the modulation of HIV-1 infection in ALFQ-treated MDM. Primary human monocytes were differentiated into MDM following in vitro culture with M-CSF media. At day 3, cultures were transfected with either APOBEC3A siRNA (A3A siRNA) or with scrambled siRNA (Control siRNA) for 24 h. At day 4 post-culture, the transfected cultures were treated with ALFQ for 24 h. MDM cultures that were neither transfected nor treated with ALFQ, or not transfected but treated with ALFQ at day 4 post-culture served as controls. (a) Cells from 3 donors were harvested, lysed and the gene expression levels of APOBEC3A were analyzed by RT-PCR. Bar graph shows fold-change (mean ± SEM) in the expression levels of APOBEC3A gene in relation to the untreated (no adjuvant) MDM controls. Data was analyzed using One-way ANOVA. (b) Cropped Western blotting (the upper band at 31kDA represents APOBEC3A) and (c) densitometry analyses show a decrease in APOBEC3A protein in the lysates of A3A siRNA cultures when compared to lysates of Control siRNA cultures. The original blot is presented in Supplementary Figs. 1–5. (d) Plots shows the percentage of intracellular p24 by flow cytometry at day 5 post-infection with HIV-1 in 3 donors (described in a) either not transfected or transfected with siRNA before treatment with ALFQ. Values in the upper right quadrant(s) of the plots represent the percentage of HIV-1-infected MDM under each treatment. (e) Scatter dot plot represent the data from (d) for triplicate wells from 3 donors and show the percentage of HIV-1-infected MDM (mean ± s.d.) in untreated MDM (black circles), MDM treated with ALFQ (magenta circles), A3AsiRNA-transfected-ALFQ-treated MDM (orange circles), and scrambled siRNA-transfected-ALFQ-treated MDM (purple circles) of triplicate wells from the 3 donors. ****P < 0.0001, One-way ANOVA.