Figure 2.

Quantitation of reverse transcriptase (RT) products using digital droplet PCR (ddPCR). MDMs from seven donors in four independent experiments were infected with R5-pseudotyped HIV-1_NL4-3. At 24 h p.i., medium was changed and 5 µM MVC was added. Cells were harvested at the indicated time points and HIV-1 RT products were quantitated in cell lysates by ddPCR as detailed in Section 2.7. (A) Scheme of the binding sites of the set of primers used for detection of early and late RT products as well as 2-long terminal repeat (LTR) circles. Cellular RPP30 was used as a housekeeping gene for normalization. (B) Copy numbers of early (5′-LTR; blue line) and late RT (Gag; red line) products or 2-LTR circles (black line) were normalized to the copy numbers of the housekeeping gene at different time points p.i. for each donor, and these values were subsequently normalized to the highest absolute number detected. Each symbol represents the mean of all the donors. Error bars represent SEM. Refer to Figure S2 (Supplementary Materials) to see individual infectivity data for all donors in the parallel experiment.