Anti α amylase antibody

Three clonal cells were analyzed for their capability to differentiate into mesenchymal cells (adipogenic, osteogenic, and chondrogenic) and hepatic cell lineages as previously described12 (link), and SG epithelial differentiation was further induced. Briefly, cells at passage 6 were seeded at 1 × 10⁴ cells/cm² onto plates that had been precoated with matrigel (BD Biosciences), after which they were cultured in serum-free hepato-STIM medium (BD Biocoat^TM, Franklin Lakes, NJ, USA) supplemented with recombinant EGF (BD Biocoat^TM), 2 mM L-glutamine, and 1% penicillin/streptomycin at 37 °C for 3 days. At the end of differentiation, cells were immunostained with anti-AQP5 antibody (Alomone Labs, Jerusalem, Israel) and anti-α-amylase antibody (Santa Cruz, CA, USA) to confirm that they were SG acinar cells. Each differentiation was further confirmed by mRNA expression of acinar cell markers as shown in Table S4.

Saliva samples obtained and were centrifuged (6000 rpm for 15 min) and stored (−70 °C) at 120 days post-RI. Saliva amylase activity was measured using salivary α-amylase assay kit (Salimetrics LLC, State College, PA, USA). Epidermal growth factor (EGF) contents were measured using an enzyme-linked immunosorbent assay (ELISA) kit (Quantikine; R&D systems, Minneapolis, MN, USA). All the experiments were performed according to the protocol. Western blotting was also conducted using an anti-α-amylase antibody (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) [5 (link)].