Oli-neuM Cell Culture and Immunoblot

Typically, 2.75 × 10⁵ Oli-neuM cells were seeded in 6-well plates in GM media and cells were grown to 70% confluence. Treatments, unless otherwise specified, were performed as indicated in the text, as previously described. For immunoblot analyses, the following antibodies were used: Cell Signaling Technology (Danvers, MA, USA): anti-phospho-p44/42 MAPK (Erk1/2Thr202/Thr204: #9101, 1:5000), anti-p44/42 MAPK (Erk1/2: #1240, 1:3000), anti-Phospho AKT (Ser473: #4060, 1:2500); anti-AKT (pan: #4691, 1:2000). Sigma-Aldrich: anti-actin (2066, 1:2000). AbD Serotec (Bio-Rad Laboratories, Hercules, CA, USA): anti-MBP (MCA409S, 1:200). Proteintech^® (Proteintech Group, Rosemont, IL, USA): anti-RXRγ (11129-1-AP,1:600). GeneTex (GeneTex, Inc., Irvine, CA, USA): anti-EGFR (GTX132810, 1:1000). Cell extract (CE) preparation and immunoblot analyses were performed as previously described [8 (link)]; briefly, bands signal intensity was estimated using ImageJ software (version 1.8.0), and data were plotted using GraphPad Prism 7.0 (GrahPad Software, San Diego, CA, USA) as the fold change versus vehicle, arbitrarily set to 1.