Anti β actin antibody ab8226

The following reagents were used: α-minimum essential medium, (α-MEM; 135-15175, WAKO, Osaka, Japan); fetal bovine serum (FBS; 10270, Gibco), anti-Arp3 antibody (sc-48344, 1:100, Santa Cruz Biotechnology); anti-β-actin antibody (ab8226, 1:100, Abcam); anti-non-muscle myosin heavy chain IIA antibody (PRB-440P-100, 1:100, BioLegend, Dedham, MA); anti-cortactin antibody [5-180, 1:100, Millipore (Upstate)]; anti-paxillin antibody (610051, 1:100, BD Biosciences); anti-vinculin antibody (V9264, 1:100, Sigma-Aldrich); anti-zyxin antibody (sc-6438, 1:100, Santa Cruz Biotechnology); rhodamine-phalloidin (R415, Molecular Probes, Thermo Fisher Scientific); fluorescein isothiocyanate-phalloidin (P5282, Sigma-Aldrich); DAPI (D1306, Molecular Probes, Thermo Fisher Scientific, CA); recombinant mouse receptor activator of nuclear factor kappa-Β (sRANK) ligand (315-11, PeproTech, NJ); fluorescein isothiocyanate- and tetramethylrhodamine isothiocyanate-conjugated secondary antibodies (715-095-151, 711-095-152 and 712-096-153, Jackson ImmunoResearch, PA); CK-666 (182515, Calbiochem, Merck Millipore, Darmstadt, Germany) and ML-7 (475880, Calbiochem).

All reagents and materials were purchased in Australia. PAR‐1 activating peptide (AP1; TFLLR‐NH₂) and PAR‐2 activating peptide (AP2; SLIGKV‐NH₂) were purchased from Auspep (Parkville, Vic., Australia). Fura‐2 acetoxymethyl ester was obtained from Thermo Fisher (Newstead, Qld, Australia). Antibodies were purchased from the following vendors: anticytokeratin (high molecular weight; 4βE12; Dako, Campbellfield, Vic., Australia), anti‐TAGLN antibody (HPA019467; Sigma‐Aldrich, Castle Hill, NSW, Australia), anti‐β‐actin antibody (ab8226; Abcam, Melbourne, Vic., Australia), anti‐αSMA (SP171; Sigma‐Aldrich), antivimentin (antiVEM, PA5‐27231; Thermo Fisher), anti‐VEGF (500‐P10‐50; Lonza, Mount Waverley, Vic., Australia), rabbit IgG isotype control and secondary antibodies (Thermo Fisher). DAPI counterstaining compound and CyQuant cell proliferation assay were purchased from Thermo Fisher. The protease inhibitor cocktail and other chemical reagents were purchased from Sigma‐Aldrich, except when specified. The Envision peroxidase system and Fast Red Substrate System were purchased from Dako. All cell culture media and reagents were purchased from Thermo Fisher, Australia, except for fetal bovine serum (FBS), which was from Sigma‐Aldrich.

Total protein was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis using 10% or 15% gels, followed by immunoblotting with the following antibodies. Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (AC002) was purchased from ABclonal Technology (Woburn, MA, USA); anti-PLCβ3 antibody (14247) was purchased from Cell Signaling Technology (Danvers, MA, USA); anti-β-actin antibody (ab8226) and anti-histone H3 antibody (ab1791) were purchased from Abcam (Cambridge, UK); anti-ace tubulin antibody (66200-1-lg) was purchased from Proteintech Group (Rosemont, IL, USA); anti-γ-catenin antibody was purchased from BD Biosciences (Franklin Lakes, NJ, USA). The GAPDH and β-actin content were analyzed as the loading control.