Es 3B and S3). We decided to use this cell line because of a smaller ratio between HIV transcripts and *mRNA in these cells compared to those in J-Lat 9.2 cells (Figure 2D, bars 3 and 4). The ratio between LTRand Rev-containing transcripts increased with the decreased levels of viral expression consistent with the rational equation (see Supplemental text 2), and revealed that the upstream transcri
M the two homologous alleles is possible. Nevertheless, these results are in a good agreement with the findings obtained by Northern blotting (Figures 1B and 1D). Taken together, the *mRNAs represent the host-viral chimeric transcripts, which contain sequences of the 5'LTR and terminate at its pA signals. In addition, RT-DqPCR revealed that levels of *mRNA exceed those of viral mRNA in unstimulate
M the two homologous alleles is possible. Nevertheless, these results are in a good agreement with the findings obtained by Northern blotting (Figures 1B and 1D). Taken together, the *mRNAs represent the host-viral chimeric transcripts, which contain sequences of the 5'LTR and terminate at its pA signals. In addition, RT-DqPCR revealed that levels of *mRNA exceed those of viral mRNA in unstimulate
Pelleted in a conical tube and washed with cold phosphate-buffered saline. Sonication and immunoprecipitation were performed using Chromatin Immunoprecipitation (ChIP) Assay Kit (Upstate) according to the manufacturer's instructions. Antibodies used are presented in Table S2. As negative control, normal rabbit or mouse serum (Sigma-Aldrich) was used. Appropriate primer pairs (Table S1) were used t
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
Ranscription units (Han et al., 2004; Lewinski et al., 2005). In the present work, we extended this knowledge by a detailed analysis of how TI impacts viral transcription in latent cells. Although TI occurs regardless of the orientation of the two promoters, we studied TI phenomenon in two J-Lat cell lines with the same orientation of the host and viral promoters, which allowed us to detect and an
Ranscription units (Han et al., 2004; Lewinski et al., 2005). In the present work, we extended this knowledge by a detailed analysis of how TI impacts viral transcription in latent cells. Although TI occurs regardless of the orientation of the two promoters, we studied TI phenomenon in two J-Lat cell lines with the same orientation of the host and viral promoters, which allowed us to detect and an
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