Fig. 3.
ACTB limits development of ASCL1+ cells in vitro. (A,B) Mouse OE explants (`e') cultured for 8 hours in ACTB (10 ng/ml) and/or FST (200 ng/ml). White arrowheads indicate ASCL1+ cells. Bar chart shows percentage of cells that are ASCL1+ (mean ± s.e.m.; *, P≤0.05 DT). (C) TgN1-2G+/– explants were cultured for a total of 30 hours, with ACTB (20 ng/ml) or GDF11 (20 ng/ml) added for the final 18 hours in vitro. GFP+ (Neurog1+) immediate neuronal precursors (INPs) were analyzed as described (Wu et al., 2003) and are plotted as mean (± s.e.m.) per 15,000 μm2 explant. *, P≤0.05 by DT. (D) Neurog1 ISH in ActβB–/– and control OE. Bar chart shows mean ± s.e.m. (E) OE explants were cultured for 6 hours, then ACTB or GDF11 (20 ng/ml) was added for 2 hours. MG132 (10 μM) or control (0.1% DMSO in medium) was added 30 minutes prior to ACTB or GDF11 addition. Analysis as in A. (F) Schematic showing actions of ACTβB and GDF11 on the ORN lineage. BL, basal lamina. Scale bars: 20 μm.