Anti itgb3

Whole-cell lysates were prepared using NP-40 lysis buffer. Protein concentrations were determined using the Bradford Assay (Thermo Fisher Scientific). Twenty milligrams of each protein lysate were loaded onto 4–20% precast polyacrylamide gels (Bio-Rad, Hercules, CA, USA), separated by electrophoresis, and electrotransferred to Sequi-Blot PVDF Membranes (Bio-Rad). Primary antibodies for Western blotting were anti-HOXB13 (Cat# PA5-78327; Thermo Fisher Scientific), anti-ITGAV (Cat# PA5-86575; Thermo Fisher Scientific), anti-ITGB5 (Cat# PA5-17260; Thermo Fisher Scientific), anti-ITGB1 (Cat# ab179471; Abcam, Cambridge, MA, USA) anti-ITGB3 (Cat# ab179473; Abcam), anti-ACTB (Cat# A5316; Sigma-Aldrich), anti-CCL2, clone 2D8 (Cat# MABN712; Merck Millipore, Burlington, MA, USA), and anti-IBSP (Cat# 5468; Cell Signaling Technology). Primary antibodies were detected by using appropriate horseradish peroxidase-conjugated secondary antibodies. Signals were then detected with an Enhanced Chemiluminescence System (GE Healthcare, Piscataway, NJ, USA) and blots were scanned using the Image Reader LAS-4000 (Fuji Film, Tokyo, Japan). Signal intensity was quantified using ImageJ software (RRID: SCR_003070; National Institutes of Health, Bethesda, MD, USA).

A western blot analysis was carried out as previously described^{31 (link)}, with the following primary antibodies: anti-G9A (Biovision; Bioptics, Tucson, USA), anti-GAPDH (Abcam; Cambridge, MA, USA), anti-SP1 (Abcam), anti-p-SP1 (Abcam), anti-Flag (Sigma-Aldrich; St. Louis, MO, USA), anti-ERK1 (Cell Signaling Technology, Danvers, MA, USA), anti-phospho-ERK1/2(Cell Signaling Technology), anti-H3 (Cell Signaling Technology), anti-H3K9me2 (Cell Signaling Technology), anti-ITGB3 (Abcam), anti-GR (Abcam), and anti-P300 (Abcam).