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
Effect of Tat on transcription of the provirus. In agreement with the results in Figure 4, expression of Tat alone exerted minimal stimulatory effect on the 5'LTR (Figures 7A-7C, Tat). However, in combination with TNF-, Tat expression activated viral transcription greatly (70-80 of GFP-expressing cells) in all three cell lines (Figures 7A-7C, Tat + TNF-). We further investigated the ability of PM
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
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
K of the latent provirus in J-Lat 9.2 cells is TI of the 5'LTR caused by the transcription of the host PP5* gene. This interfering transcription results in the lack of Sp1 and the presence of the elongating RNAPII at the 5'LTR. Moreover, TI can be partially rescued by TNF- stimulation, as documented by the appearance of Sp1 and initiating RNAPII at the 5'LTR. The slight enrichment of the RNAPII in
M et al.Author ManuscriptAuthor ManuscriptAuthor ManuscriptGautam et al.DFZWPage13.11.11.10.9.6.2.2.1.1.0.0.0.DFZV16.13.14.12.12.11.10.10.0.ACKNOWLEDGEMENTSWe thank R. Plishka, A. Peach and T. Lewis for determining plasma viral RNA loads and K. Rice, R. Engel, R. Petros and S. Fong for diligently assisting in the maintenance of animals and assisting with procedures. We also thank R. Schwartz, Vacc
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
Y i is given by:Verguet Cost Effectiveness and Resource Allocation 2013, 11:1 http://www.resource-allocation.com/content/11/1/Page 3 ofTable 1 Cost parameters for the male circumcision interventionIntervention Initial investment per circumcision facility CF Initial training per circumciser CT Salary of each circumciser CC Salary of health care workers/counselors per circumciser Cost of supplies pe