Figure 5
From: N-terminal Slit2 inhibits HIV-1 replication by regulating the actin cytoskeleton
Slit2N mediates HIV-1 inhibition by binding to the Robo1 receptor. (A) MT4 cells transfected with non-targeting (NT) or Robo- siRNA by nucleofection (Amaxa biosystems) were analyzed for Robo1 expression by Western blotting. GAPDH was used as a loading control. (B) The NT and Robo1 siRNA-transfected cells were pre-treated with Slit2N (3nM) or control, and left uninfected or infected with HIV-1 IIIB for 48 hours. The supernatants were analyzed for HIV-1 p24 levels by ELISA. *P < 0.05 versus the mock-infected cells transfected with NT-siRNA. (C) MT4 cells were stimulated with HIV-1 gp120 for various periods of time in the presence and absence of Slit2N. The lysates were immunoprecipitated (IP) with anti-Robo1 antibody. The immune complexes were then immunoblotted with β-actin antibody. The same blot was reprobed with Robo1 antibody. TCL- total cell lysate. IgG- antibody control. Results are representative of 3 separate experiments.