Ab5320

For immunofluorescence, cells cultured on 9 mm coverslips and nanofiber inserts were used. After designated culture periods, the cells were fixed for 20 min with 4 % paraformaldehyde, washed three times with PBS, and stored at 4 °C until use. The cells were blocked with 10 % normal goat serum (NGS) and 0.1 % Triton X-100 in PBS for 1 h at room temperature. Cells were washed with PBS and were incubated overnight at 4 °C with the following primary antibodies in the blocking solution; anti-NG2 (1:500, Millipore, #AB5320), anti-NG2 (1:500, Santa Cruz Biotechnology, #sc-33666), anti-MBP (1:500, Abcam, #AB7349), anti-Olig2 (1:500, Merck Millipore, #MABN50), anti-GFAP (1:2000, Dako, #Z033429), anti-GFAP (1:2000, Abcam, #AB4674), anti-Tuj1 (1:1000, Dako, #G7121), anti-Iba1 (1:500, Wako, #019–19741), anti-cleaved-caspase-3 (1:200, Cell Signaling, #9664S), anti-Ki67 (1:500, Thermo Fisher Scientific, #MA514520), anti-BCAS1 (1:500, Synapse Systems, #445-003). The cells were washed with PBS and incubated for 1hr, RT with the appropriate Alexa Fluor 488-, 594-, and 680-conjugated secondary antibodies (Thermo Fisher Scientific). Subsequently, the cells were counterstained with 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI, Sigma Aldrich, #D9542) for 10 min at RT and mounted onto glass slides. The cells were imaged using a Zeiss LSM800 confocal microscope.

HIV-1 gp120-IIIB (Cat # 11784) was provided by the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH. For immunoblotting, we used the following antibodies: anti-NG2 (1:1000, Millipore: AB5320), anti-PDGFRα (1:1000, Santa Cruz: sc-338), anti-Olig2 (1:2000, Millipore: MABN50), anti-MBP (1:5000, Covance: SMI 94 and SMI 99), anti-proteolipid protein (PLP) (1:1000, Thermo: PA3-150), and anti-β-actin (1:1000, Santa Cruz: sc-1616-R).