Control igg antibody

The RIP assay was performed as previously described.^[31] Briefly, the MagnaRIP RNA‐Binding Protein Immunoprecipitation Kit (Millipore, MA, USA) was used according to the manufacturer's instructions. Corresponding cell lysates were incubated with beads coated with 5 µg of control IgG antibody (Beyotime, Shanghai, China), anti‐APAF1 antibody (ab234436, Abcam, MA, USA), or anti‐IGF2BP1 antibody (22803‐1‐AP, Proteintech, IL, USA) with rotation at 4 °C overnight. Next, total RNA was extracted for the detection of ABL expression by qRT‐PCR.

Monoclonal rabbit anti-SDC-1, anti-VCAM-1, anti-MyD88, anti-TLR4, anti-p-IκBα-Ser36 antibodies and monoclonal mouse anti-CD31 antibodies were purchased from Abcam (Cambridge, UK). Monoclonal rabbit anti-vascular endothelial- (VE-) cadherin and anti-NF-κB/p65 antibodies were bought from Cell Signaling Technology (Boston, MA, USA). Tetramethylrhodamine- (TRITC-) phalloidin (which binds to F-actin) was from Solarbio (Beijing, China), and 4′,6-diamidino-2-phenylindole (DAPI) was from SouthernBiotech (Birmingham, USA). Anti-HMGB1 antibody was from Wanleibio (Shenyang, China), and control IgG antibody was from Beyotime (Shanghai, China). Glycyrrhizin (GLY; an HMGB1 inhibitor) was obtained from JK Scientific (Beijing, China). TAK242 (a TLR4 inhibitor), ST2825 (a MyD88 inhibitor), and SN50 (an inhibitor of NF-κB/p65 translocation) were purchased from MedChemExpress (NJ, USA). Fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) was purchased from Solarbio (Beijing, China). A human SDC-1 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Abcam (Cambridge, UK), and a mouse SDC-1 ELISA kit and VCAM-1 ELISA kit were purchased from Zcibio (Shanghai, China). A Total Nitric Oxide (NO) Assay Kit was purchased from Beyotime Biotechnology (Shanghai, China).