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
Ns dormant. In contrast, we demonstrated that TI could be counteracted by specific inhibition of the upstream transcription or by cooperative activation of transcription initiation and elongation from the 5'LTR by the viral and host TFs. In the latter scenario, we envision that RNAPII initiating from the viral promoter competes successfully with the RNAPII that is elongating through the 5'LTR. Bec
Ns dormant. In contrast, we demonstrated that TI could be counteracted by specific inhibition of the upstream transcription or by cooperative activation of transcription initiation and elongation from the 5'LTR by the viral and host TFs. In the latter scenario, we envision that RNAPII initiating from the viral promoter competes successfully with the RNAPII that is elongating through the 5'LTR. Bec
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
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
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
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
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