stemsep human cd8 + depletion kit  (STEMCELL Technologies Inc)

Journal: The Journal of Biological Chemistry

Article Title: Multiple NF-?B Sites in HIV-1 Subtype C Long Terminal Repeat Confer Superior Magnitude of Transcription and Thereby the Enhanced Viral Predominance *

doi: 10.1074/jbc.M112.397158

Figure Lengend Snippet: 4-κB viruses out-compete the 3-κB strains in T-cells A, schematic representation of the paired isogenic viral constructs used in the competition assays. The 22-bp sequence consisting of the F-κB site was engineered into the 3′-LTR of the Indie-C1 molecular clone (HHC) to generate the 4-κB LTR (FHHC). Replication profiles of the viral strains in vitro using CEM-CCR5 T-cells (B) or the CD8-depleted, mitogen-activated PBMC (C) from a representative subject. The cells were infected with 500 infectious units of FHHC or HHC viruses, and the secretion of p24 into the medium was monitored for several weeks as indicated. The data are presented as the mean of triplicate wells ± 1 S.D. and are representative of three independent experiments. Pairwise competition between the HHC and FHHC isogenic viruses in CEM-CCR5 cells (D) or activated PBMC (E). The paired viruses were competed against each other at different ratios of m.o.i. as schematically depicted in supplemental Fig. 4. Genomic DNA was extracted on day 10 following the viral infection and subjected to the HTA analysis. The heteroduplex band intensities are plotted as relative values compared with the monoinfections (see formula supplemental Fig. 4D).

Article Snippet: The CD8 cells were depleted from the PBMC using the StemSep Human CD8 + depletion kit (catalog no. 14662, Stem Cell Technologies, Vancouver, Canada).

Techniques: Construct, Sequencing, In Vitro, Infection