Mouse anti xbp1s

Individual retinas (8–10) from P4 - P28 tubby and wt mice were collected, homogenized and proteins were quantified (Cai et al., 2012 (link)). Soluble proteins, 30 μg, were loaded in 10% SDS-page gels and electrophoresed. After electrophoretic transfer to the nitrocellulose membrane, the following primary antibodies at 1:1000 dilution were used: phospho-eIF2α (Ser51)(119A11) rabbit mAb and phospho-Bcl2 (Ser70)(5H2) rabbit mAb (Cell signaling technology, 3597 and 2827, respectively), mouse anti-ATF6 (Imgenex, IMG-273), rabbit anti-GRP78 BiP and rabbit anti-IRE1 (phospho S724) (Abcam, ab48187), rabbit NF-κB p65 (C-20) (Santa Cruz Biotechnology, sc-372) and mouse anti-XBP-1s (Biolegend, 647502). β-actin (13E5) rabbit mAb (HRP conjugate) (Cell signaling technology, 5125) was used as a loading control. The membrane development, imaging and densitometry of the bands were performed as previously reported (Cai et al., 2012 (link); Kong et al., 2011 (link)).