Rabbit anti human bax

Rabbit anti-human Bax, ERK, JNK and p-JNK antibodies were obtained from ProteinTech Group (Chicago, IL, USA). Rabbit anti-human antibodies against p-PI3K, AKT, p-AKT, PARP, pro-caspase-3, cleaved-caspase-3, pro-caspase-9, cleaved-caspase-9, Bcl-xl, Bcl-2, Mcl-1, Bad, p38, p-p38, p-ERK, p-ATF2 and p53 were obtained from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-human antibodies against p-Bad and cytochrome C were obtained from Epitomics (Burlingame, CA, USA). Rabbit anti-human β-actin antibodies were obtained from Sigma (St Louis, MO, USA). Goat anti-rabbit and horseradish peroxidase conjugated IgG was obtained from Millipore (Bellerica, MA, USA). Immunoblotting was performed as previously described [19 (link)]. The treated samples and the control samples (25 μg) were separated by 12.5% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The proteins on the gel were then transferred to PVDF membranes. The PVDF membranes were incubated with the primary antibody (1:1000 dilutions in 2% dehydrated skim milk) at 4 °C overnight. Then, incubation at 4 °C was performed for 2 h with the secondary antibodies (goat anti-rabbit or goat anti-mouse and horseradish peroxidase conjugate, 1:5000 dilution in 2% dehydrated skim milk). The blots were detected through chemiluminescence using enhanced ECL western blotting kit.

Fucoxanthin purchased from MedChemExpress (New Jersey, USA) (purity 99.17%) was dissolved in dimethyl sulfoxide (DMSO) to make a 50 mM stock and stored at -20℃ and diluted with fresh culture medium before use. An equal volume of DMSO (final concentration < 0.1%) as added to the controls. TGF-β1 was purchased from PeproTech (Rocky Hill, NJ, USA). Primary monoclonal antibodies including rabbit anti-human Bax, Bcl-2, Caspase 3, Cleaved Caspase 3, PARP, Cleaved PARP, E-cadherin, Vimentin, β-catenin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibodies were purchased from ProteinTech Group, Inc. (Chicago, IL, USA). The Alexa Fluor® 790-conjugated donkey anti-rabbit secondary antibodies were purchased from Abcam (Cambridge, MA).