Col1a col 1

The mouse monoclonal antibodies COL1A (COL-1) (diluted 1:500, sc-59772) and LLH1 (B-5) (diluted 1:500, sc-59772; PLOD1) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The rabbit polyclonal antibodies COL4A (diluted 1:1000, ab6586) and P3H1 (diluted 1:500, ab154799) were purchased from Abcam, (Cambridge, UK). The rabbit polyclonal antibodies P3H2 (diluted 1:500, 15723-1-AP), P3H4 (diluted 1:500, 15288-1-AP), and P4HA2 (diluted 1:500, 13759-1-AP) were purchased from Proteintech (Rosemount, IL, USA). The mouse monoclonal antibody anti-α-tubulin (DM1A) (diluted 1:1000, #T9026) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The anti-mouse Ig (diluted 1:200, Alexa Fluor™ Plus 594) and anti-rabbit Ig (diluted 1:200, Alexa Fluor™ Plus 488) were purchased from Thermo Fisher Scientific.

Proteins (100 µg) were applied on Novex NuPAGE 4–12% Bis-Tris gel (Invitrogen Life Technologies) under nonreducing conditions and transferred to a 0.2 µm Hybond nitrocellulose membrane (GE Healthcare). Staining with Ponceau S and β-actin were used as loading controls. The membranes were incubated with antibodies against β-Actin (AC-74, Sigma), p21 (F-5, Santa Cruz), p53 (BP53-12, Exbio), BCl-2 (C-2, Santa Cruz), Bax (B-9, Santa Cruz), Timp-1 (D10E6, Cell Signaling), α-SMA (1A4, Sigma), cellular fibronectin (IST-9, Santa Cruz), TGF-β1 (V, Santa Cruz), Col1A (COL-1, Santa Cruz), SDHA (B-1, Sabta Cruz), SDHB (B-1, Santa Cruz), UCP2 (G-6, Santa Cruz), SIRT3 (F-10, Santa Cruz), TRAP-1 (C-8, Santa Cruz), SUNCR1 (GPR91, Abcam), and Pannexin 1 (Abcam) at 4 °C overnight. Secondary antibodies were from Sigma Aldrich. Detection was performed with Western Blotting Luminol Reagent (Santa Cruz) and SuperSignal West Femto (ThermoFisher Scientific). Blots were scanned and quantified using a PXi imaging system (Syngene, Cambridge, UK).