Ormed using NorthernMax kit (Ambion) according to the manufacturer's instructions. The probes were amplified from cDNA synthesized with MMLV reverse transcriptase (Invitrogen). PCR products were labeled with BioNick Labelling System (Invitrogen). Stimulation of J-Lat cells and detection of activated cells by flow cytometry Tat protein was expressed from pCDNA3 vector using electroporation of 107 J
Surgery. Costs include fixed costs and functional costs. Fixed costs include medical equipment and certification of trained circumcisers. Functional costs, which are variable, include oversight and promotion (management, monitoring, communication and advertising), salaries of full-time medical practitioners, surgical staff and counselors (for each medical practitioner, we allocate 1 medical assist
Clinically relevant resistance mutations in the protease gene. The most frequent RT mutations were M184V (n?11), conferring resistance to lamivudine and emtricitabine, and Y181C (n?4), G190A/S (n?4) and K103N (n?4), conferring resistance to NNRTIs. Of concern, three children (16 ; 95 CI: 3 ?40 ) had thymidine analogue mutations, associated with cross-resistance to all NRTIs. Figure 1 gives the pr
And from cells expressing Tat protein, 48 h after electroporation. qPCR was performed in the presence of SyBr Green (Sigma). Primer sequences and positions are in Table S1. Construction of stably transfected cell lines LMP vectors (Open Biosystems) expressing two microRNA-adapted shRNAs (shRNAmir) against the ER and a control shRNAmir (Table S1) were transfected into Phoenix cells with FuGENE6 rea
And from cells expressing Tat protein, 48 h after electroporation. qPCR was performed in the presence of SyBr Green (Sigma). Primer sequences and positions are in Table S1. Construction of stably transfected cell lines LMP vectors (Open Biosystems) expressing two microRNA-adapted shRNAs (shRNAmir) against the ER and a control shRNAmir (Table S1) were transfected into Phoenix cells with FuGENE6 rea
Me transcription through the provirus, since slight enrichment of elongating RNAPII in the HIV coding region of transcriptionally silent J-Lat 9.2 cells was observed using ChIPqPCR assay. However, transcription originating from the host promoter, ignoring pA sites in both LTRs and consequently splicing out the provirus together with the host intron (Han et al., 2004) is most likely less frequent t
Ly 20 activation of viral transcription (see Figure 7) already precludes detection of the upstream transcription. This finding is not surprising, as we demonstrated that there is 11-fold or 38-fold more viral mRNA than *mRNA in TNF- treated cells (Figure 2D, bars 3 and 4). Predictably, our control experiments demonstrate that the ratio between the Rev- and Env-containing transcripts was 1 in untr
Ly 20 activation of viral transcription (see Figure 7) already precludes detection of the upstream transcription. This finding is not surprising, as we demonstrated that there is 11-fold or 38-fold more viral mRNA than *mRNA in TNF- treated cells (Figure 2D, bars 3 and 4). Predictably, our control experiments demonstrate that the ratio between the Rev- and Env-containing transcripts was 1 in untr