Fig 1: TGFβR inhibition represses RGDSP-induced K7 expression. (a) hS/PCs were cultured in HA and RGDSP hydrogels with A83-01 or the vehicle control (DMSO) for 14 days and visualized with bright field microscopy. (b) dsDNA, indicative of proliferation, was resolved from simultaneous TRIzol RNA/protein extractions using the Quant-iT PicoGreen assay. (c,d,f–h) hS/PCs were cultured in HA and RGDSP hydrogels for 14 days receiving A83-01 or DMSO, and expression of KRT7 (c), FN (d), IGF2 (f), KRT7-AS (g), and AMY1A (h) was assessed with qPCR. (e) hS/PCs were cultured in HA and RGDSP hydrogels for 14 days receiving A83-01 or DMSO, and western blotting was conducted to investigate K7 and fibronectin expression. Blotting for fibronectin and K7 was performed sequentially on the same membrane (Fig. S7g–i). Duplicate measurements are presented from technical replicates extracted from separate hydrogels. (i) hS/PCs were cultured for 14 days in RGDSP receiving A83-01 or DMSO, and fluorescent microscopy indicated that expression α-amylase and β-catenin was not suppressed by treatment with A83-01. Single channel images are presented in Fig. S13. Quantification was conducted from 3 independent experiments, each with 3 technical repeats. Error bars represent SEM in all cases. One-way ANOVAs were performed on data presented in (b-h) followed by Tukey’s multiple comparison test. * indicates p < 0.05 in all cases.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for Human Salivary Amylase Alpha ELISA Kit (Chemiluminescence)