Donkey anti rabbit igg h l secondary antibody

Cells were grown on 24-well plates during 24 h before hydrostatic pressure treatment (40 mmHg, 24 h). Then, they were fixed with a paraformaldehyde solution (4%) before incubation with anti-laminin antibody (1:400, Abcam, UK) overnight at 4°C. After extensive washing, plates were incubated for 2 h at room temperature with green-fluorescent donkey anti-rabbit IgG (H+L) secondary antibody (1:500, Invitrogen, USA). 4',6-diamidino-2-phenylindole was used to stain the cell nuclei (blue). Finally, cells were observed under a fluorescence microscope (IX71, Olympus, Tokyo, Japan) and images were analyzed with Image-Pro Plus software (Image-Pro Plus 6.0, Media Cybernetics, Inc., Silver Spring, MD, USA).

Anti-GRP78 (Cat# ab21685, RRID: AB_2119834), anti-ATF6 (Cat# ab83504, RRID: AB_2058904), anti-p-IRE1 (Cat# ab124945, RRID: AB_11001365), anti-XBP1 (Cat# ab220783), anti-eIF2α (Cat# ab26197, RRID: AB_2096478), anti-p-eIF2α (Cat# ab32157, RRID: AB_732117), anti-LC3B (Cat# ab192890, RRID: AB_2827794) and anti-Caspase-3 (Cat# ab13847, RRID: AB_443014) antibodies were purchased from Abcam. Anti-CHOP (Cat# 2895, RRID: AB_2089254) antibody was from Cell Signaling Technology, and anti-LC3B (Cat# L7543, RRID: AB_796155) antibody, 4-phenylbutyric acid (4-PBA), tunicamycin (Tm), cholesterol, L-α-phosphatidylcholine, L-glutamine, polybrene, and puromycin were obtained from Sigma. Chemically Defined Lipids and Opti-MEM medium were purchased from Gibco. TurboFect was from Thermo Fisher Scientific and anti-β-actin antibody was obtained from Tianjin Sanjian Biotech. HRP-conjugated goat anti-rabbit and anti-mouse secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology, and donkey anti-rabbit IgG (H + L) secondary antibody and Alexa Fluor 594 were from Invitrogen. 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) was from Beyotime Biotechnology.