FIG. 4.
Production of infectious HIV-1 is not affected by CD63 knockdown. MDM were nucleofected with scrambled or CD63-specific siRNA and infected with HIV-1BaL as described in the text. (A) CD63 expression levels were assessed by Western blot analysis at 6 and 10 dpi. The clathrin heavy chain (CHC) was analyzed as a loading control. The percentage of knockdown is shown in the relevant lanes. (B to D) Cell-free supernatants from control and CD63 knockdown (KD) HIV-1BaL-infected MDM were assayed for p24 levels (B) or infectivity (C), and the number of infectious units per particle were calculated (D). Data from experiments with five different donors were averaged and are represented as relative units after normalizing with respect to control values of p24, infectivity, and infectious units per particle at 6 dpi. For panels B through D, bars represent the average relative units, and error bars represent standard errors of the means. (E) Env and Gag (p24 and pr55) expression levels in control and CD63 knockdown HIV-infected MDM at 6 and 10 dpi were assessed by Western blotting.