Acy 738

N-Acetyl-L-aspartic acid (Sigma-Aldrich, Burlington, MA, USA) was dissolved in water containing 8% NaOH and used at a final concentration indicated in the text. Clobetasol (Prestw-781) and Gefitinib (Prestw-1270) were purchased from Prestwick Chemical Library^®, Illkirch-Graffenstaden, France and used at a final concentration of 10 µM or 1 µM, respectively, as previously described [30 (link)]. The HDAC inhibitor ACY-738 (HY-19327, MedChemExpress, Monmouth Junction, NJ, USA) was dissolved in 100% DMSO at a final concentration of 10 mM and used at a concentration of 10 µM, according to [31 (link)]. After which, 0.5% DMSO max was added to NAA or vehicle treatments in all experiments, while Clobetasol, Gefitinib, or ACY-738 were used in parallel experiments as controls. Drug treatments were administrated in DM for 48 h to cells left to grow in GM for 24 h. Culturing and time of drug treatments have been established previously based on the timing and concentration for optimal MBP expression in Oli-neuM [29 (link),30 (link)]. Engagement tests were performed as previously described in growth chambers containing polystyrene (PS) microfibers of 2–4 µm. For microfiber engagement analysis, the cells were treated for a minimum of 72 h, according to previously established protocols [28 (link),29 (link)].

Primary antibodies used for immunocytochemistry (ICC) and WB included mouse anti-aaTub (1:10,000; Sigma, St. Louis, MO, USA; Cat #T6793), rabbit anti-ARL13B antibody (1:5000; Proteintech, Rosemont, IL, USA; Cat #17711-1-AP), mouse anti-TUJ1 (1:2000; R&D Systems, Minneapolis, MN, USA; Cat #NL1195V), chicken anti-Ki-67 (1:5000; Encor, Gainesville, FL, USA; Cat# CPCA-Ki67), chicken anti-GFAP (1:1000; Encor, Gainesville, FL, USA; Cat# CPCA-GFAP), mouse anti-GAPDH (1:10,000; Encor, Gainesville, FL, USA; Cat# MCA-1D4), chicken anti-GFP (1:5000; Abcam, Cambridge, UK; #ab13970), rabbit anti-HDAC6 (1:1000; Sigma, St. Louis, MO, USA; Cat #H2287), rabbit anti-HDAC6 (1:1000; Cell Signaling, Danvers, MA, USA; Cat #7558S), mouse anti-KIF3A (BD Biosciences, San Jose, CA, USA; Cat #611508), and rabbit anti-PCM1 (1:1000; Bethyl Laboratories, Montgomery, TX, USA; Cat# A301-150A). ACY-1215 (APEXbio, Houston, TX, USA; Cat #A4083) and ACY-738 (MedChemExpress, Monmouth Junction, NJ, USA; Cat #1375465-91-0) were dissolved in 100% DMSO (Fisher Scientific, Waltham, MA, USA; Cat # D128-500) to produce a stock concentration of 10 mM. cDNA vectors pCMV-EGFP or pCMV-mycEGFP:HDAC6[#NM_001321225.2] were designed and obtained from Vectorbuilder (Vectorbuilder.com (accessed on 29 March 2021)).