Lation than the 5'LTR, suggesting that the latter LTR is much stronger subjected to TI. Even if there is transcription from host gene `reading through' both LTRs in some latent cells, this does not diminish the impact of TI on the 5'LTR.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptCell Host Microbe. Author manuscript; available in PMC 2014 November 03.Lenasi et al.Page
Lation than the 5'LTR, suggesting that the latter LTR is much stronger subjected to TI. Even if there is transcription from host gene `reading through' both LTRs in some latent cells, this does not diminish the impact of TI on the 5'LTR.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptCell Host Microbe. Author manuscript; available in PMC 2014 November 03.Lenasi et al.Page
Lation than the 5'LTR, suggesting that the latter LTR is much stronger subjected to TI. Even if there is transcription from host gene `reading through' both LTRs in some latent cells, this does not diminish the impact of TI on the 5'LTR.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptCell Host Microbe. Author manuscript; available in PMC 2014 November 03.Lenasi et al.Page
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
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