Figure 8: Nuclear YAP is reduced in mSCs of Dy2j/2j mice and increasing nuclear YAP rescues short internode phenotype. | Nature Communications

Figure 8: Nuclear YAP is reduced in mSCs of Dy2j/2j mice and increasing nuclear YAP rescues short internode phenotype.

From: Optimal myelin elongation relies on YAP activation by axonal growth and inhibition by Crb3/Hippo pathway

Figure 8

(a) Teased nerve fibres of Dy2j/2j mice or wild-type littermates (Dy+/+) at 15 dPN immunostained for myelin (MBP, red), paranodes (Caspr, green) and nuclei (DAPI, blue). (b,c) Quantification of total YAP (c) and phosphorylated YAP (P-YAP) (b), normalized to actin, in sciatic nerves from Dy+/+ (4 animals) and Dy2j/2j (five animals) littermates. (d) YAP immunostaining (green) in mSCs nuclei (DAPI,blue) at 7 dPN in Dy2j/2j and Dy+/+ littermates. (e) Quantification of YAP immunostaining in nuclei of Dy2j/2j (black circle) and Dy+/+ mSCs (white circle) at 7, 14 and 21 dPN. The number of nuclei analysed between 38 and 120 (three animals). (f) A Dy2j/2j mSC infected with Crb3 shRNA virus (lower panels, DsRed2) has more YAP (green) in its nucleus (DAPI, blue, red arrows) than a non-infected (NI) cell in the same nerve (upper panels, white arrows). Virus injection at 3–4 dPN and sacrifice at 30 dPN. (g) Quantification of nuclear YAP immunostaining in Dy2j/2j mSCs, not-infected (NI, 87 nuclei) or infected with control (Contsh, 18 nuclei) or Crb3 (Crb3sh, 13 nuclei) shRNA viruses. (h) mSCs infected with Control shRNA virus in a control littermate nerve (Dy+/+ Cont sh, GFP, top) or in a Dy2j/2j mutant mouse nerve (Dy2j/2j-Cont sh, GFP, middle) or infected with Crb3 shRNA virus in a Dy2j/2j mutant mouse nerve (Dy2j/2j-Crb3 sh, DsRed2, bottom). Mouse pups were injected at 3–4 dPN and sacrificed at 30 dPN. (i) Quantification of internodal lengths of cells described in h. The reduction of internodal length in cells of Dy2j/2j mice (Dy2j/2j-Cont sh, 41 cells) versus wild-type littermates (Dy+/+ Cont sh, 62 cells) is partially compensated when Dy2j/2j mouse cells are silenced for Crb3 (Dy2j/2j-Crb3 sh, 76 cells). a.u., arbitrary units. Scale bars, 50 μm (a), 15 μm (d), 20 μm (f) and 100 μm (h). Statistical two-tailed Student’s t-tests: *P<0.05; **P<0.01; ***P<0.001. Error bars show s.d in b, c and s.e.m in e, g and i.

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