Journal: Virology

Article Title: miR-132 enhances HIV-1 replication

doi: 10.1016/j.virol.2012.12.016

Figure Lengend Snippet: A) Resting CD4+ T cells were isolated from four healthy blood donors, and a portion of the cells was activated with PHA. Activated CD4+ T cells were harvested two days later, and total RNA was isolated for use in microRNA microarray analysis. Columns represent fold-change in miR-132 expression following activation. B) Resting CD4+ T cells were isolated from five healthy blood donors, and a portion of the cells was activated with anti-CD3/28 beads. Two days later, total RNA was isolated and quantitative real-time PCR was performed to measure miR-132 levels relative to U6 snRNA; values presented were normalized relative to miR-132 levels in the resting cells of Donor 5. MiR-132 levels were found to be significantly higher following CD4+ T cell activation (Student’s t-test, p < 0.02). C) Relative miR-132 levels were measured as above in resting or PHA-activated PBMCs from HIV-positive, viremic donors; no significant difference in miR-132 levels was observed between activated cells from young versus elderly donors (Student’s t-test, p > 0.05).

Article Snippet: Resting CD4 + T cells were isolated from healthy donor blood (Gulf Coast Regional Blood Center, Houston, TX) using the RosetteSep human CD4 + T cell enrichment cocktail (StemCell); activated cells were removed using CD30 Microbeads (Miltenyi Biotec).

Techniques: Isolation, Microarray, Expressing, Activation Assay, Real-time Polymerase Chain Reaction