Figure 4.

Effect of RGC-32 deletion on GFAP and vimentin expression in vitro in mouse astrocytes. WT and RGC-32 KO mouse astrocytes were isolated from the brains of 2-day-old pups and grown in DMEM/F-12 medium supplemented with 10% fetal bovine serum for 2 weeks, plated on a slide culture chamber, and then stained with anti-GFAP antibody. WT astrocytes show a mature morphology (A), whereas RGC-32 KO astrocytes have a radial glia like morphology (B). Original magnification: x40. Scale bars: 10 µm. The expression of GFAP and vimentin was analyzed by Western blotting and normalized to β-tubulin. Significantly higher levels of GFAP protein are seen in WT astrocytes than in RGC-32 KO astrocytes (C), whereas vimentin levels are higher in KO astrocytes (D). Results in (C, D) are expressed as mean ± SEM (n=3), and a representative blot is shown for each protein. * = p<0.05; ** = p<0.01.