Ab36447

The ADPs (passage 3) were grown on a four-well chambered cell culture slide. After fixation with 4% paraformaldehyde and washing with PBS, the cells were incubated with primary antibody against CD146 (dilution 1:100; ab210072), PDGFR-β (dilution 1:100; ab69506), NG2 (dilution 1:100; ab50009), CD45 (dilution 1:100; ab10558), CD34 (dilution 1:100; ab81289), CD31 (dilution 1:100; ab119339), CD73 (dilution 1:100; ab175396), CD90 (dilution 1:100; ab225), CD105 (dilution 1:100; ab11414), Nestin (dilution 1:100; ab11306), and MAP2 (dilution 1:100; ab36447) (all from Abcam, see Supplemental Table 1) in PBS/0.1%Tween 20 and 10% normal goat serum overnight at 4°C. The cells were washed with PBS and incubated with secondary antibodies: goat anti-rabbit IgG Alexa Fluor 594 (dilution 1:500; A-11012), goat anti-rabbit IgG Alexa Fluor 488 (dilution 1:500; A-11008), and goat anti-mouse IgG Alexa Fluor 488 (dilution 1:500; A-11029) (all from Thermo Fisher Scientific, Waltham, MA, USA) in PBS for 45 min at room temperature. After washing in PBS, the cells were mounted in antifade reagent with 4’,6-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific) for 1 min and visualized with an inverted fluorescence microscope Axio Observer A1 (Carl Zeiss, Jena, Germany).