Immunostaining of Brain Cells

Immunostaining of brain sections or dissociated neural cells was performed as described previously (Kim et al. 2004 (link); Ka et al. 2016b (link)). The following primary antibodies were used: Mouse anti-NeuN (EMD Millipore, MAB337), rat anti-neural cell adhesion molecule L1 (EMD Millipore, MAB5272), rabbit anti-TBR1 (Abcam, ab31940), rabbit anti-CUX1 (Santa Cruz, sc-13024), chicken anti-MAP2 (Abcam, ab5392), rabbit anti-GFAP (Abcam, ab7260), rabbit anti-Ki67 (Cell Signaling, #9129), rabbit anti-phospho-Histone H3 (Cell Signaling, #9701S), mouse anti-BrdU (BD Biosciences, 555627), chicken anti-Tbr2 (EMD Millipore, ab15894), rabbit anti-BLBP (Abcam, ab32423), mouse anti-Nestin (PhosphoSolutions, 1435-NES), rabbit anti-ARL13B (Abcam, ab83879), rabbit anti-Acetyl-α-Tubulin (Cell Signaling, #5335), chicken anti-GFP (Thermo Fisher, A10262), rabbit anti-GFP (Thermo Fisher, A11122) and mouse anti-MACF1 (Santa Cruz, sc-377532). Alexa Fluor568-Phalloidin (Thermo Fisher, A12380) was used to stain polymerized actin. Appropriate secondary antibodies conjugated with Alexa Fluor dyes (Thermo Fisher) were used to detect primary antibodies. The purpose of using each antibody is listed in Table1.

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Ka M., Moffat J.J, & Kim W.Y. (2021). MACF1 involved in the 1p34.2p34.3 microdeletion syndrome is essential in cortical progenitor polarity and brain integrity. Cellular and molecular neurobiology, 42(7), 2187-2204.

Publication 2021

MAB337 rabbit anti-CUX1 rabbit anti-Ki67 rabbit anti-phospho-Histone H3 mouse anti-BrdU rabbit anti-Acetyl-α-Tubulin chicken anti-GFP rabbit anti-GFP Alexa Fluor568-Phalloidin Alexa Fluor dyes

Antibodies Antibody Brain Brdu Chicken Dyes Gfap Histone h3 Map2 Mouse Nestin Neural cell adhesion molecule l1 Neural cells Phalloidin Polymerized actin Rabbit Stain α tubulin

Corresponding Organization : Kent State University

Other organizations : Korea Institute of Toxicology, University of California, San Francisco

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Immunostaining patterns of various neural markers (NeuN, neural cell adhesion molecule L1, TBR1, CUX1, MAP2, GFAP, Ki67, phospho-Histone H3, BrdU, Tbr2, BLBP, Nestin, ARL13B, Acetyl-α-Tubulin, MACF1), and polymerized actin (stained with Alexa Fluor568-Phalloidin)

control variables

None explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

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