Anti xbp1s antibody

Whole-protein isolations were performed using RIPA lysis buffer (Santa Cruz Biotechnology, TX, USA). Proteins were separated by SDS-PAGE using TGX stain-free gels (AnykD and 4–15% gels, Bio-Rad) and transferred to a PVDF membrane (Bio-Rad) by wet electroblotting (Bio-Rad). Total loaded protein was used for normalization of target protein expression (Image Lab 6.0.1, Bio-Rad). The PVDF membranes were blocked for 15 minutes with EveryBlot (Bio-Rad) at room temperature, followed by incubation overnight with a recombinant anti-LC3B antibody [EPR18709] (1:1000 at 4°C, ab192890, Abcam, UK), anti-CysLT1 antibody (1:500 at RT, ab151484, Abcam), anti-ATF4 antibody (1:1000 at RT, Cell Signaling, MA, USA), anti-ATF6 antibody (1:1000 at RT, Cell Signaling) or anti-XBP1s antibody (1:500 at RT, BioLegend, CA, USA) diluted in EveryBlot. Antibodies against LC3B, CysLTR1, ATF4 and ATF6 were labeled using an anti-rabbit antibody conjugated to HRP (Agilent, CA, USA), and anti-XBP1s was labeled using an anti-mouse antibody conjugated to HRP (Agilent) diluted in EveryBlot. The antibodies were visualized using Clarity Western ECL Substrate (Bio-Rad) and the ChemiDoc XRS + system (Bio-Rad).