Collagen 3a

Proteins were extracted from each group of left ventricular tissues with lysis buffer supplemented with protease inhibitor and phenylmethylsulfonyl fluoride (PMSF). Proteins (20 μg /sample) were separated by electrophoresis with 10% SDS/PAGE gels. After electrophoresis, proteins were transferred to nitrocellulose membranes. The blots were blocked with 5% non-fat milk in Tris-buffered saline with Tween-20 (TBS-T) for 1 h at room temperature, and then incubated with primary antibodies against mouse LOX-1 (a gift from Dr. Sawamura; 1:2,000, v/v, dilution), fibronectin (Abcam; 1:2,000, v/v, dilution), collagen-1a (Santa Cruz, Dallas, TX, USA; 1:1,000, v/v, dilution), collagen-3a (Santa Cruz; 1:1,000, v/v, dilution), collagen-4a (Santa Cruz; 1:1,000, v/v, dilution), p22^phox (Santa Cruz; 1:1,000, v/v, dilution), gp91^phox (Abcam; 1:2,000, v/v, dilution) or β-actin (Santa Cruz; 1:2,000, v/v, dilution) in TBS-T at 4°C overnight on a shaker. After washing with TBS-T three times, the blots were incubated with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz) for 1 h at room temperature. The blots were scanned with a ChemiDOC XRS system (Bio-Rad, Hercules, CA, USA) following exposure to Luminol Reagents (Beyotime, Shanghai, China) for 3 min.

hUCB-MSCs were obtained from MEDIPOST Co. Ltd (Seoul, Korea). Fetal bovine serum was bought from BioWhittaker Inc. (Walkersville, MO, USA). AA, A23187, bisindolylmaleimide I, BrdU, Indomethacin, LY294002, mitomycin C, Nordihydroguaiaretic acid (NDGA), rapamycin, 1-Aminobenzotriazole (1-ABT), and SB203580 were aquired from the Sigma Chemical Company (St Louis, MO, USA). Phospho-Akt^ser473, phospho-Akt^thr308, Akt1/2/3, β-Actin, collagen1A, collagen3A, collagen5A, fibronectin, phospho-p38, p38, phospho-JNK, JNK, p-ERK1/2, ERK, lamin A/C, MMP-12, pan-cadherin, PKCα, PKCɛ, PKCθ, PKCζ, PKC, Phospho-PKCζ, and Sp1 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Phospho-PKC, phospho-mTOR^ser2481 (mTORC2), phospho-mTOR^ser2448 (mTORC1), and mTOR were purchased from Cell Signaling (Beverly, MA, USA). The Akt inhibitor I was aquired from Calbiochem (La Jolla, CA, USA). The CD34, GPR40, phospho-Sp1, and MT3-MMP antibodies were obtained from Abcam (Cambridge, MA, USA). Mithramycin A was purchased from Tocris (Bristol, UK). Zymogram gels were bought from Novex (San Diego, CA, USA). Horseradish peroxidase-conjugated goat anti-mouse and goat anti-rabbit IgG were obtained from Jackson Immunoresearch (West Grove, PA, USA). All other reagents were used as received and were of the highest purity commercially available.

Imatinib mesylate (IMT) was procured from Sigma Aldrich, USA. Cerulein was obtained from Ana Spec, France. TGF-β1 was procured from Bio-legend, USA. The antibodies viz. β-actin (Catalogue no. sc-47778), Collagen1a (Catalogue no. sc-393573), Collagen3a (Catalogue no. sc-271249), α-SMA (Catalogue no. sc-53142), CTGF (Catalogue no. sc-373936), TGF-β1 (Catalogue no. sc-146), DDR2 (Catalogue no. sc-81707), pSmad2/3 (Catalogue no. #8828), Smad2/3 (Catalogue no. #8685), pDDR1 (Catalogue no. #14531), DDR1 (Catalogue no. #5583), pNF-κB (Catalogue no. #3033), NF-κB (Catalogue no. #8242) were procured from Santa Cruz Biotechnology Ltd and Cell Signaling Technology, USA. The ELISA kits of TGF-β, IL-1β, IL-6 and TNF-α (Catalogue no. TGF-β: 88–8350, IL-1β: 88–7013, IL-6: 88- 7064, and TNF-α: 88–7324) were obtained from eBioscience, USA.