Fig. 4

Triple activation of GR, ERα, and RARα enhanced neuronal differentiation in KCN69n and IMR32 cells. A Immunofluorescence staining after ligand treatment. The upper two panels show KCN69n and the lower two panels IMR32 cells. Green = SCG2; red = Phalloidin; blue = DAPI. Scale bars represent 20 μm. B Western blot analysis of neural differentiation markers in KCN-GR + EV, KCN-GR + ERα, IMR32-GR + EV, and IMR32-GR + ERα cells following incubation with ligands. Molecular weight markers shown to the left and β-actin was used as a loading control. Cells were treated with 20 nM E2, 200 nM DEX, 1 μM ATRA, or the combination of either 20 nM E2 + 200 nM DEX, or 20 nM E2 + 200 nM DEX + 1 μM ATRA for seven days. Experiments were performed at least three times