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. 2013 Jun 11;8(6):e65818. doi: 10.1371/journal.pone.0065818

Figure 5. Endogenous Shh acts as a niche factor.

Figure 5

(A) Nsps expresses high levels of Shh ligand that co-localizes with Nestin, a NSC marker. (B) Western blots analysis shows that Shh is expressed in nsps even in control conditions. The Shh ligand is endogenously expressed in E18.5 tectum; cerebellum is shown as a positive control. (C) Ptc1 immunostaining images of nsps revealed increased expression of the receptor in the presence of Shh. Image is shown in 3-D projection resulting of a clockwise 135° rotation in the Z-axis. (D) Representative images of ALPA of C3H-10T1/2 cells, treated with fresh conditioned serum-free media obtained from 2 day cultured nsp populations, conditioned media plus Cyc, or Pur (used as positive control) in comparison to nsp media without growth factors used as control media. (E) Percentage of ALPA positive cells in response to indicated treatments. Bar, 20 µm. *** p<0.0001. W/O GF: without growth factors, E: EGF, F: FGF-2. CM: conditioned media.