Figure 1.

METH and HIV-1 gp120 IIIb induce autophagy in astrocytes. LC3II was analyzed using the western blot and quantified by AlphaEase FC software (Alpha Innotech, San Leandro, CA, USA), which are shown at the bottom of each panel (a–f). Results are shown as mean±S.E. from three separate experiments. *P<0.05. (a) SVGA cells were exposed to different doses of METH for 24 h, (b) SVGA cells were exposed to 1 mM of METH at varying time periods, (c) SVGA cells were exposed to different doses of HIV-1 gp120 IIIb for 24 h, (d) SVGA cells were exposed to 400 pM gp120 IIIb at varying time periods, (e) SVGA cells were exposed to 1 mM of METH, 400 pM of gp120 IIIb, or both for 24 h, (f) human primary astrocytes were treated with METH and gp120 IIIb as indicated, (g) SVGA cells were exposed to bafilomycin A1 1 h before treatment of METH, gp120 IIIb, or both for 24 h. (h) LC3II punctate dots in METH- and gp120- treated SVGA cells. SVGA cells were treated with 1 mM of METH, 400 pM of gp120 IIIb, or both for 24 h. Cells were fixed with ice-cold methanol: acetone (1 : 1), immunostained with anti-LC3II antibody, and examined by confocal microscopy (scale bar, 20 μm). (i) Electron microscopy images showing the ultrastructure of METH- and gp120-treated SVGA cells. Arrows in the electron micrograph denote presence of autophagosomes (scale bar, 2 μm). Immunostaining and microscopic images are representatives of at least three independent experiments