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. Author manuscript; available in PMC: 2015 Nov 19.
Published in final edited form as: Nat Commun. 2014 Nov 19;5:5455. doi: 10.1038/ncomms6455

Figure 2. Inhibition of Wnt signalling restored neuronal differentiation of Cdo-deficient cells.

Figure 2

(a) Luciferase assays of P19 cells expressing either pSuper or Cdo shRNA treated either DMSO or XAV939. (b) Fluorescence microscopy of pSuper or Cdo shRNA expressing P19 cells at ITS1 immunostained with β-tubulin III antibodies. Size bar = 100 µm. (c) Quantification of the representative experiment shown in b. P19 clusters that possess various numbers of β-tubulin III-positive cells were counted and plotted as percentile. More than 6 fields and at least 10 clusters/field were counted. Data are means±s.d. (n = 6) (d) Immunoblot analysis of cell lysates from P19/pSuper or P19/shCdo treated with XAV939 or the vehicle DMSO in the ITS medium for 24 h. (e) qPCR with RNAs isolated from primary neurospheres of Cdo+/+ or Cdo−/− NPCs treated with either DMSO or 4 µM XAV939 for 24 h in the absence of bFGF. (f) Confocal microscopy of Cdo+/+ and Cdo−/− NPCs treated with DMSO or 4 µM of XAV939 for 24 h on the differentiation day 1, followed by immunostaining for β-tubulin III. Size bar = 50 µm. (g) Quantification of the relative signal intensity of β-tubulin III from data shown in f. The mean values of at least 10 fields. Values are means ±s.d. from the determinants of triplicates. All experiments in this figure were repeated three times with similar results. *P<0.05, **P<0.01, ***P< 0.005. Statistical significance was tested by a Student t-test.