Blots was subsequently probed with primary antibodies, including rabbit polyclonal anti-Capn4 antibody (1 1000, LifeSpan Biosciences, Seattle, WA, USA), anti-MMP2, anti-MMP9, anti-Snail, anti-Vimentin, anti-E-cadherin, anti-H-cadherin, anti-β-catenin antibodies (1 1000, Cell Signaling Technology, Danvers, MA, USA), anti-NF-κB/p65 (SC-109, 1: 2000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).
The blots were subsequently probed with anti-phospho-ser32/36-IκB-α anti-phospho-ser32/36-IκB-α anti-phospho-ser32/36-IκB-α anti-phospho-ser32/36-IκB-α anti-phospho-ser32/36-IκB-α
After gel electrophoresis, blots were subsequently probed with primary antibodies (anti- IL-6, 1∶1000 #ab6672, antIL-17-1∶30003000 #ab40663, anti-TGF-β 1∶1000 #ab66043 Abcam Inc; Cambridge, MA).
Blots were subsequently probed with horseradish peroxidase-conjugated anti-mouse or anti-rabbit secondary antibodies (Amersham Biosciences, Buckinghamshire, England) diluted 1∶5000 in 1× PBS, 0.1% tween-20 and 5% fat-free dry milk.
Blots were subsequently probed with Clean-Blot IP-horseradish peroxidase conjugate (Thermo Scientific).
Blots were subsequently probed with anti α-tubulin (Sigma, UK) to show equal sample loading.
Blots were subsequently probed with anti- α-tubulin (Sigma, Poole, UK) to show equal sample loading.
These fractions were subsequently probed for LDH1.
These blots were subsequently re-probed with an antibody against total ERK and p38 (New England Biolabs, Inc .. Bands were visualised as before.
Electrophoresis and Western blotting was subsequently carried out.
