Fig 10. Inhibition of MVB and vacuole formation reduced HIV-1 sequestration and virus spread to lymphocytes.
(A) Polarized tonsil cells were transfected with control siRNA or siRNAs against Hrs and rabankyrin-5. After 72 h, expression of Hrs and rabankyrin-5 was examined by Western blot. The mean density of protein bands is shown under the blot. β-Actin was detected to confirm equal loading. Immunoblots were performed at least twice and a representative figure is shown. (B, C) Tonsil epithelial cells transfected with control siRNA or siRNAs specific to Hrs or rabankyrin-5 at 72 h after transfection were exposed to HIV-192UG029 or HIV-1SF170. After 3 days, one set of siRNA-transfected cells were examined for intracellular HIV-1 (upper panels). The next set of siRNA-transfected cells were cocultured with activated CD4+ T lymphocytes (lower panels). Four hours later, lymphocytes were collected and grown for 4 days for HIV-192UG029 and 10 days for HIV-1SF170. HIV-1 infection was examined by ELISA p24. Data represent one of three independent experiments and are shown as mean ± SEM of triplicate values. ***P < 0.0001 and ****P < 0.00001, compared with the control siRNAs.