Fig. 5

Minimal interactions between HIV replication and CycT1 levels in memory CD4 T cells. a Memory CD4 T cells were purified from peripheral blood, then uninfected or infected with HIV (R5 strain NSN-SX) in IL2 medium for 2 days. Cells were washed and cultured with CD3+CD28 mabs and IL2 for 6 days. Cells were then stained for intracellular p24 in conjunction with either CycT1, phosphorylated-CDK9 (Thr186), CD25, CD69, CD38, HLA.DR, Ki67, p21, beta7 integrin, CD4, CD62L, or HLA.ABC. Shown are sample dotplots of uninfected and HIV-infected bystander p24- and infected p24+ cells, and mean ± sem expression of proteins in uninfected and infected bystander p24- and infected p24+ cells (*p < 0.05 comparing p24- to p24+ cells, N = 3–5). b Reduction of HIV replication concomitant with CycT1 reduction in memory CD4 T cells by raltegravir. HIV-infected memory CD4 T cells (5 days post-infection) were treated with AZT, raltegravir, or saquinavir for 3 days, then stained for p24, CycT1, CD25, and CD69. Shown are mean ± sem p24 and CycT1 levels gated on CD69+CD25+ cells (*p < 0.05, N = 3). c Comparison of P-TEFb protein levels between GFP- and GFP+ J-Lat6.3 cell lines by western blot. Shown are western blots of CycT1, CycT2, pCDK9, and total CDK9 in non-sorted or sorted (following 24 h PMA treatment) GFP- or GFP+ J-Lat cells