Monoclonal rabbit anti pakt

Western blot for protein expression was performed as described previously^{19 (link)}. HUVECs seeded in 6 well plates were processed 30 min, 1 h, 4 h and 6 h after shockwave treatment. The blots were probed with monoclonal rabbit anti-pAKT, monoclonal rabbit anti-AKT, monoclonal mouse anti-pERK (all Cell Signaling Technology, Cambridge, UK), polyclonal rabbit anti-ERK (Santa Cruz Biotechnology, Dallas, US, MA) and mouse β-actin (Sigma Aldrich, St. Louis, MO) antibody.

Cells were lysed with lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1.5 mM MgCl₂, 5 mM EDTA, and 1% Triton^™ X-100) containing protease and phosphatase inhibitor cocktails. Samples prepared as described above were separated by SDS-PAGE and then transferred onto Polyvinylidene difluoride membranes (Merck Millipore, Billerica, MA, USA). After blocking, the membranes were incubated with the following primary antibodies: polyclonal rabbit anti-FGFR4 (ab41948; Abcam, Cambridge, UK), monoclonal rabbit anti-ERK1/2 (#4695; Cell Signaling Technology, Danvers, MA, USA), monoclonal rabbit anti-pERK1/2 (#4370; Cell Signaling Technology), monoclonal rabbit anti-AKT (#4691; Cell Signaling Technology), monoclonal rabbit anti-pAKT (#4060; Cell Signaling Technology), monoclonal mouse anti-STAT3 (#9139; Cell Signaling Technology), polyclonal rabbit anti-pSTAT3 (#9131; Cell Signaling Technology), monoclonal rabbit anti-MT1-MMP (ab51074; Abcam), and monoclonal mouse anti-β-actin (A5316; Sigma-Aldrich). Membranes were then incubated with the appropriate peroxidase-conjugated secondary antibodies (Cell Signaling Technology), washed, and developed with ECL^™ Prime reagents (GE Healthcare, Piscataway, NJ, USA). Image densitometry was performed with ImageJ software (National Institutes of Health, Bethesda, MD, USA). Full blot images with molecular weight markers are shown in Figure S10.