Rabbit anti baf45d

Immunofluorescence (IF) assay was performed according to our previous protocol (Liu et al., 2017 (link)). The samples were first incubated with rabbit anti-BAF45D (1:100, Proteintech, Chicago, IL, United States) together with mouse anti-NEUN (1:100, Millipore, Belecula, CA, United States), mouse anti-MBP (1:500, Abcam) and mouse anti-GFAP (1:100, Proteintech, Chicago, IL, United States) overnight at 4°C. After washing in PBS, the samples were then incubated with Alexa Flour-488 anti-mouse (1:500) and Alexa Fluor-594 anti-rabbit (1:500) secondary antibodies. Nuclei were counterstained with DAPI. The IF assay for cultured cells was performed in accordance with our previous report (Liu et al., 2007 (link)) and positive staining was visualized with a NIKON Eclipse 80i fluorescence microscope and a NIKON Eclipse Ti-S inverted fluorescence microscope.

The lysates of fresh tissues and the H9-derived cells were subjected to IB assay according to the previous protocol (Liu et al., 2012 (link); Tripathi and Mishra, 2012 (link)). Then the proteins were detected using indicated antibodies. Antibodies used for IB are as follows: mouse anti-GFAP (1:500, Proteintech, Chicago, IL, USA), mouse anti-NEUN (1:500, Millipore, Belecula, CA, USA), rabbit anti-BAF45D (1:500, Proteintech, Chicago, IL, USA), rabbit anti-GATA6 (1:100, Abcam, New Territories, HK), mouse anti-OCT4 (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-PAX6 (1:500, Millipore, Belecula, CA, USA) and rabbit anti-GAPDH (1:2000, Proteintech, Chicago, IL, USA) antibodies. More descriptions of the antibodies used for IB were shown in Table S1.

Lysates were prepared from fresh tissues and cells and then subjected to immunoblotting (IB) in accordance with a previously described protocol (Liu et al., 2017 (link)). Proteins were then detected using a range of antibodies: mouse anti-GFAP (1:2000, Proteintech, Chicago, IL, United States), mouse anti-NEUN (1:500, Millipore, Belecula, CA, United States), rabbit anti-BAF45D (1:500, Proteintech, Chicago, IL, United States), mouse anti-beta-III-tubulin (1:1000, Arigo) and rabbit anti-GAPDH (1:2000, Proteintech, Chicago, IL, United States). Specific details of the antibodies used for IF and IB are given in Supplementary Table S2.