Genesnap acquisition software

The lysates were obtained by lysing cells in lysis buffer containing 50 mM Tris, pH7.4, 150 mM NaCl, 0.25% sodium deoxycholate, 1% NP-40, 0.1% SDS, 1 mM PMSF, and complete protease inhibitor cocktail (Roche, Mannheim, Germany). Equal amounts of total protein were subjected to 10% SDS-PAGE and blotted onto PVDF membranes (Pall, Pensacola, FL). Western blotting was performed using rabbit anti-human COL3A1 antibody (Bioss, Beijing, China). The blotting membranes were scanned using GeneSnap acquisition software (Syngene, Cambridge, UK) and band densities were quantified with the GeneTool program (Syngene, Synoptics). GAPDH were used as internal control.

Phages were concentrated using polyethylene glycol (PEG; Sigma-Aldrich, USA) precipitation as previously described [26 ]. After mixing with PEG8000, the phage suspension was centrifuged at 11,000×g for 25 minutes at 4°C and the pellet was re-suspended in SM buffer (Storage Media; 100 mM NaCl, 10 mM MgSO₄.7H₂O, 50 mM Tris-HCl, pH7.5). DNaseI (14 mg/ml) and RNaseA (30 mg/ml) (Thermo Fisher Scientific, USA) were then added, and the mixture was incubated at 37°C for 1 hour to eliminate bacterial nucleic acids. Following phenol/chloroform extraction, the aqueous phase was precipitated using absolute ethanol. The precipitated phage DNA was washed, dissolved in sterile deionized water, and stored at -20°C until use.
For restriction enzyme analysis, the isolated phage DNA was digested with BstBI or MluI restriction endonucleases using the manufacturer’s recommended conditions. Digested DNA was separated by agarose gel electrophoresis, and images were captured using GeneSnap acquisition software (Syngene, UK).