Mpared GFP expression of parental and J-Lat shER cells using fluorescence activated cell sorting (FACS). As expected, no GFP-expressing cells could be detected in untreated cells (Figures 6C-6F, mock). About 20 of cells turned GFP-positive in TNF--treated J-Lat 9.2 cells (Figure 6C, TNF-). Importantly, the inhibition of PP5 transcription synergized with TNF- to increase further the number of GFP-
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
T the level of gene expression act cooperatively to establish and maintain HIV latency. In conclusion, our study reveals that TI represents a key mechanism that antagonizes proviral gene expression to promote the latency of HIV. Furthermore, it demonstrates several means that could be used to counteract TI for reactivating latent HIV. Future mechanistic studies linking TI and other transcriptional
T the level of gene expression act cooperatively to establish and maintain HIV latency. In conclusion, our study reveals that TI represents a key mechanism that antagonizes proviral gene expression to promote the latency of HIV. Furthermore, it demonstrates several means that could be used to counteract TI for reactivating latent HIV. Future mechanistic studies linking TI and other transcriptional
Ed cells. Furthermore, TNF- stimulation led to the appearance of Sp1 and initiating RNAPII on the 5'LTR and additional enrichments of initiating and elongating RNAPII on the 3'LTR. These effects are most likely achieved through the activation of NF-B, which stimulates several steps of viral transcription including the PIC formation, transcription initiation and transcription elongation. Thus, our
Ed cells. Furthermore, TNF- stimulation led to the appearance of Sp1 and initiating RNAPII on the 5'LTR and additional enrichments of initiating and elongating RNAPII on the 3'LTR. These effects are most likely achieved through the activation of NF-B, which stimulates several steps of viral transcription including the PIC formation, transcription initiation and transcription elongation. Thus, our
Vels of viral particles, we speculate that average levels of host-viral chimeric transcripts in these cells exceed those in the two J-Lat cell lines. These results thus suggest that TI plays a key role in primary CD4+ T cells. In J-Lat 9.2 and 15.4 cells, the host-viral chimeric transcripts terminated at the pA site in the 5'LTR, but transcripts initiating at the host promoter and terminating at t
Vels of viral particles, we speculate that average levels of host-viral chimeric transcripts in these cells exceed those in the two J-Lat cell lines. These results thus suggest that TI plays a key role in primary CD4+ T cells. In J-Lat 9.2 and 15.4 cells, the host-viral chimeric transcripts terminated at the pA site in the 5'LTR, but transcripts initiating at the host promoter and terminating at t