Skip to main content
. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Genesis. 2013 Aug 16;51(10):10.1002/dvg.22415. doi: 10.1002/dvg.22415

FIG. 2.

FIG. 2

Tet-regulatable f-BRAF expression in vitro. Ad5-Cre mediated recombination results in GFP expression in cerebellar astrocytes (a) and NSCs (b), which is silenced following the addition of 1µg/ml doxycycline for 3 days (line 21). (c) GFP immunostaining reveals expression of the transgene (green) in Ad5-Cre-infected neurospheres. f-BRAF expression in astrocytes (d) and NSCs (e) results in increased MAPK activation (Thr-202/Tyr-204 phosphorylation). α-tubulin is included as an internal loading control. RNA RT-PCR confirms the presence of the transgene (d, e; right panel). β-actin is included as an internal control. (f) There was no change in astrocyte proliferation following f-BRAF expression. (g) f-BRAF expression results in increased NSC proliferation. Ad5-LacZ infected cells serve as controls. (h) Limiting dilution assay shows a higher frequency of secondary neurosphere formation in f-BRAF (Cre)-expressing NSCs. Error bars denote mean ± SD. Scale bar, 100µm. (*) P< 0.01. ns, not significant.