Figure 2. HIV-1 Envelope Induces mTOR Signaling Pathway Activation.
(A) The upper panel shows an immunoblot of DC lysates with PathScan I cocktail recognizing phosphorylated mTOR pathway proteins after 15 min of HIV-1 exposure. Arrows indicate bands corresponding to Erk and S6 phosphorylated proteins. Experiment shown is representative of five independent experiments.
(B) Lysates of DC untreated or treated with HIV-1 or with HIV-1-ΔEnv (500 ng p24 each) for 15 min and immunoblotted with PathScan I cocktail. Arrow indicates S6 phosphorylated protein. Data are representative of three independent experiments.
(C) Immunoblot with PathScan I cocktail of lysates of DCs untreated or treated for 15 min with HIV-1 or HIV-1-F522Y. Arrows indicate bands corresponding to Akt, Erk, and S6 phosphorylated proteins. Loading was controlled with anti-actin (lower panel). Data are representative of two experiments.
(D) PathScan I immunoblotting on lysates of 5 × 105 DCs untreated or treated for 15 min with mouse Ig (10 μg/ml), rabbit Ig (10 μg/ml), mouse anti-CD4 (10 μg/ml), rabbit anti-DC-Sign (H-200 at 10 μg/ml), rgp120 (5 μg/ml), rantes (10 nM), and HIV-1 (1 μg p24). Phosphorylated Erk and S6 migrating bands are indicated by arrows. Similar results were obtained in three independent experiments.
(E) Lysates of DCs untreated or treated with HIV-1, HIV-1-ΔEnv, or HIV-1-F522Y (500 ng p24 each) for 15 min and immunoblotted with anti-phospho2448-mTOR. Arrow indicates phosphorylated mTOR protein. Equivalent loading was controlled by immunoblotting with anti-mTOR (lower panel). Data are representative of three independent experiments.
(F) Lysates of iDCs untreated or incubated for 12 hr with HIV-1 (1 μg p24), Ig isotype (10 μg/ml), rgp120 (5 μg/ml), and anti-CD4 (10 μg/ml) were immunoblotted with anti-LC3 (upper panels). Loading was controlled with anti-actin (lower panels). Data are representative of two experiments.