Figure 1. YAP/TAZ are largely nuclear in proliferating, differentiating and mature myelinating SCs.

YAP/TAZ localization was investigated in sciatic nerve cryosections (A, B) or teased fibers (C, D). SC nuclei are marked by Sox10 (red) or DAPI (blue). (A) YAP/TAZ (green) are in SC nuclei throughout development, from E16.5 to P60. (B) YAP phosphorylated on Ser 112 (p-YAP, green) is largely absent from SC nuclei at P60. (C) Co-localization of YAP (green) in the nuclei of myelinating SCs marked by Krox20 (white) at P60. Arrows point to mSC nuclei that contain YAP and Krox20; arrowheads point to non-myelinating SC nuclei, which lack Krox20 and YAP. (D) YAP/TAZ (green) are present in the nucleus of a mSC, associated with a large axon, but is absent from the nucleus of a non-myelinating SC, marked by GFAP (red), at P60. Arrow points to myelinating SC nucleus; arrowhead points to non-myelinating SC nucleus. (E) Diagram summarizing the developmental stages in which YAP/TAZ are present or absent from the SC nucleus. The following figure supplements are available for Figure 1.

DOI: http://dx.doi.org/10.7554/eLife.20982.002

Figure 1—figure supplement 1. Selection of specific antibodies for immunohistochemical analysis of YAP and TAZ.

(A) Western blotting of WT P23 sciatic nerve lysates, showing three different antibodies specifically binding to both YAP/TAZ, only YAP, and only to phosphorylated YAP. (B–D) These antibodies were further tested in transverse sciatic nerve sections of adult WT, Yap cKO, Taz cKO and Yap/Taz cDKO mice. Sections were co-stained with anti-Sox10 (red) to mark SC nuclei, and DAPI to label all cellular nuclei. (B) An antibody specifically recognizing both YAP and TAZ. Its immunoreactivity completely disappears only in cDKO SCs. (C) An antibody selectively binding to YAP. Its immunoreactivity is eliminated in YAP cKO SCs and Yap/Taz cDKO SCs, but not in Taz cKO SCs. Arrows in WT denote examples of SC nuclei exhibiting YAP immunoreactivity. (D) An antibody (S112) selectively binding to phosphorylated YAP (p-YAP). Its immunoreactivity is largely cytoplasmic in WT, and is eliminated in Yap cKO SCs and Yap/Taz cDKO SCs, but not in Taz cKO.

DOI: http://dx.doi.org/10.7554/eLife.20982.003