Fig. 3

Tracking and quantification of HIV/STAT5B and HIV/BACH2 chimeric transcripts in different hematopoietic cell subsets obtained from HIV patients under cART. a Top panel, agarose gel electrophoresis of the RT-PCR products (using the primers indicated in Fig. 1c) obtained from the cDNA of the indicated cell subsets purified by FACS sorting from two HIV-1 patients (ID-96, ID-77). For each cell subsets RT-PCR for GAPDH was also performed as positive control of amplification (bottom panels). In both patients, a specific band of 500 bp was obtained from the cDNA extracted from the purified Treg and Tcm cell subsets. In ID_77 two different bands appeared in the agarose gel due to splice variants of the STAT5B gene. In PBMC the band is detectable only when 10-fold more cDNA was loaded in the RT-PCR. The band was not detected in all the other cell types, including Tscm, T effector memory (Tem), and others. b Top panel, agarose gel electrophoresis of the RT-PCR products obtained from the cDNA of the indicated cell subsets purified by magnetic cell isolation in four HIV-1 patients (ID-76, ID-56, ID-25, ID-63). RT-PCR for GAPDH was also performed as positive control of amplification (bottom panel). M: molecular size marker. c, d Histograms show the average of the relative levels of the copies of the HIV/STAT5B (c) and of the HIV/BACH2 chimeric transcript (d) vs. HPRT copies measured by dd-PCR in Treg and Teff cells obtained from PBMC of six and three HIV-infected patients, respectively. e, f Histograms indicate the relative levels of the copies of STAT5B (e) and BACH2 (f) vs. HPRT copies measured by dd-PCR in Treg and Teff cells obtained from PBMC of six and three HIV-infected patients, respectively. Significance was determined by Mann–Whitney tests (**p < 0.01), SEM is indicated by the error bar