Figure 7. Env trafficking to the TRE regulates particle incorporation in primary MDMs.
(A) MDMs were prepared from human donor monocytes and infected with HIV-1BAL. Cells were stained for HIV-1 Env and MICAL-L1 on day 5 post-infection. Region of interest shown as inset on right demonstrates colocalization of Env with this TRE marker. (B) siRNA-mediated depletion of MICAL-L1 or EHD1 was carried out in MDMs prior to infection with HIV-1BAL, using two distinct siRNAs and compared with Scr siRNA. Cell lysates and particles were harvested on day 7 following infection and examined by Western blot. Note depletion of gp41 and gp120 bands in siRNA lanes. (C) Quantitation of Env incorporation as assessed by gp41/p24 ratio in four separate experiments employing depletion of MICAL-L1 (M-siRNA) or EHD1 (E-siRNA). Significance addressed using one-way ANOVA as before. ***, P<0.001.