Quantitative Western Blot Analysis of Cellular Fractions

Whole cell, cytosolic and nuclear homogenates from cells as well as the mouse cortices and hippocampi were prepared as previously described (Marwarha et al. 2014a (link), Marwarha et al. 2017b , Marwarha et al. 2017c , Marwarha et al. 2010a (link), Marwarha et al. 2010b (link)). Proteins (10–40μg) were resolved on SDS-PAGE gels followed by transfer to a polyvinylidene difluoride membrane (BioRad, Hercules, CA) and incubation with the monoclonal antibodies listed in Table 4. The origin, source, the dilutions of the respective antibodies used for this study is compiled in Table 4. β-actin was used as a gel loading control for whole cell and cytosolic homogenates, whereas Histone H3 was used as a gel loading control for nuclear homogenates. The blots were developed with enhanced chemiluminescence (Clarity^TM Western ECL blotting substrate, Bio-Rad, Hercules, CA) and imaged using a LiCOR Odyssey Fc imaging system.