Fig. 6

Estrogen receptor α protein expression and effect of β-estradiol on neural progenitor cells proliferation. Germ like cells, hNP cells, DIV 14 neuronal cells were seeded in 8 well slide for 24 h at density of 30,000 cells per well and fixed for estrogen receptor α protein immunostaining. Germ like cells, hNP cells and DIV 14 neuronal cells were also seeded in 60 mm dish and collected at 90% confluent density for western blot experiment. In proliferation assay, hNP cells were seeded onto 96 well plates at a density of 25,000 cells/well, medium was changed with β-estradiol every other day until end of DIV 3 and DIV 14. All data are from 3 separate experiments using independent cultures (n = 18 wells total). *Concentration is significant different from control group (P < 0.05, one-way ANOVA). a: Germ like cell estrogen receptor α staining; b: hNP cell estrogen receptor α staining c: Western blot analysis of IMR90, GLCs, hNP and DIV 14 neuron estrogen receptor α expression. β-actin was used for normalizing the loading of samples. d: DIV 3 cell proliferation assy. d: DIV 14 cell proliferation assy. Scale bars = 50 μm