Fig 8. MAPK signaling pathway participates in proliferation and hepatic differentiation of hAECs.

(A) Amniotic cells were seeded in 24-wells plate and incubated in IMDM medium supplemented with 10% FBS (C) or hepatic differentiation medium (HD), during indicated times. At each time cells were fixed and Ki-67 expression (green) was detected using Alexa-488 conjugated secondary antibody. Representative micrographs from hAECs at different HD times at 10X are shown. The nuclei were stained with DAPI (blue). Percentage of positive cells (fluorescence intensity) for Ki-67 is shown in the upper right graph. Scale bar: 60 μm. Representative results from three replicates are shown. (B) Amniotic cells were incubated during 30 days with IMDM medium 10% FBS (C) or hepatic differentiation medium (HD), with or without PD98059, as indicated. Extracts from cells were prepared as previously described and loaded in a 12% SDS-PAGE. Albumin expression was determined by Western-blot. Loading controls were performed by immunoblotting the same membranes with anti-GAPDH. Bands densitometry is shown in lower panels. Molecular weight (kDa) is indicated at the right of the blot. Representative results from three replicates are shown. ***p<0.001 vs. control; ###p<0.001 vs. respective control.