Fig. 3

Role of Pin1 in HIV integration in CCL19-treated resting CD4⁺ T cells. Resting CD4⁺ T cells were cultured with CCL19 or activated with PHA-IL2 for 2 days prior to infection with HIV NL4-3. Immunoprecipitated proteins were analysed by SDS-PAGE and immunoblot with anti-HIV integrase antibody. a Co-IP of integrase and Pin1 was determined by IP with anti-Pin1 antibodies and probing for integrase (32 kDa) and Pin1 (18 kDa) in the presence and absence of the JNK inhibitor (SP600125, 10 µM) in HIV-infected CCL19-treated (upper panel) and PHA-IL2 activated (lower panel) CD4⁺ T cells. b PHA-IL2 activated and CCL19-treated resting CD4⁺ T cells were transfected with Pin1-specific or scrambled control siRNA, infected 2 days later with HIV, and 5 h later examined for Pin1 expression by immunoblot with antibodies to Pin1 or the GAPDH loading control (lower panel). The effects of Pin1 siRNA inhibition on HIV nuclear entry and integration were determined by real time PCR quantification of HIV 2-LTR circles (left panels) or Alu-LTR (right panels) respectively at day 4 post-infection. HIV-infected PHA-IL2 activated (grey columns) or CCL19-treated (open columns) CD4⁺ T cells are shown. Each column represents the mean of three donors. Individual donors are shown as symbols. *p < 0.05; **p < 0.01