Figure 3
ING5 activates mitogenic pathways to promote self-renewal. (a) The sphere formation rates of iPB-ctr and iPB-ING5 overexpressing cells at three successive passages in the absence of EGF and FGF treatment. (n=3, **P<0.01, ***P<0.001). (b) DIC images of spheres from the tertiary sphere passage in iPB cells. Scale bar=400 μm. (c) Sphere formation rate under treatment with protein kinase inhibitors (n=3, *P<0.05). (d) Protein and phosphorylated protein levels of effectors in the PI3K and MEK pathways. Cells were treated with PX-866 at 1 μM and PD184352 at 2 μM for 48 h. (e) Immunofluorescence of Nestin and Tubb3 in differentiated iPB control (Left panels) or iPB-ING5 cells (Right panels) treated with 1 μM PX-866 or 2 μM PD184352. Scale bar=200 μm. (f) Flow cytometry analysis of the CD133 positive population in PX-866 (1 μM) and PD184352 (2 μM) treated BTIC 189 cells, gated by isotype control.
