Fig. 2

Effects of AsA on collagen maturation and differentiation in MSCs. a Proliferation of cells in collagen-coated dishes (orange line) and non-coated polystyrene dish (gray line). Data for the latter are the same data as those in Fig. 1a. b IHC of Col1. Col1 antibody: green, DAPI: blue. c Sirius Red/Fast Green staining for collagen. 100% confluent cells were used for staining. d Quantification of total collagen. After staining, the amounts of collagen and non-collagen proteins were inferred by measuring the absorption at 540 nm (Sirius Red) and 605 nm (Fast Green). e Adipogenic differentiation staining. After culturing the cells to 100% confluence, the culture medium was replaced with adipogenic differentiation medium. After 7 days of culturing, lipids were stained with Oil Red O. In the bottom row, nuclei are counterstained with DAPI. f Quantitative evaluation of adipogenic differentiation. The relative adipogenic differentiation rate was expressed as the number of cells that underwent adipogenic differentiation divided by the total number of cells at an AAP of 0 mM