Paving the way toward a cure? Study reports new insights into role of proteins in HIV latency NatureComms
at room temperature for 1 h in the presence of 10 µg/ml polybrene . Following spinoculation, virus was removed, and fresh medium was added to the well for 4 days. Infected cells were then run and analyzed by flow cytometry ). Post-acquisition analysis was performed on a separate computer using FlowJo . The proportions of latent or productive integrations were calculated by dividing the percentage of or cells, respectively, by the total percentage of positive cells.
A dilution of this was analyzed by ddPCR as described using the same conditions described above and the following primers: LambdaR-REV2 5’-GTT TCG CTT ACG TGG CAT CAG ACG G-3’ and Late U3-FWD 5’-GCT ACA TAT AAG CAG CTG CTT TTT GCC TGT AC −3’. The following probe was used: /5YAkYel/ CTT TAT TGA GGC T + T AAG + C + AG + T + G + GG T/3IABkFQ. Nucleotides followed by a+ indicate an LNA base to improve the melting temperature of the probe.
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