DEV023820 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Table S2 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Analysis of the cell division plane in proliferative chondrocytes infected with RCAS-GFP. (A) A representative image of infected chondrocytes (green) containing a cell at telophase (arrow) with a well-defined actin-rich contractile ring (red) separating two daughter nuclei (blue). Scale bar: 25 µm. (B) Telophase θ for GFP-expressing proliferative chondrocytes is highly similar to that of wild-type uninfected proliferative chondrocytes (P=0.82).

• Supplemental Figure S2 -

  Fig. S2. Analysis of gene expression in the developing chick humerus. Sections from uninfected humeri (WT) and humeri expressing Fzd7^-C or Vangl2 were analyzed by staining with Hematoxylin and Eosin (H&E) and by in situ hybridization for Col1aI (Col I), Col2a1 (Col II), Col9a1 (Col IX) and Indian hedgehog (IHH). Arrows in Col I samples indicate signal in the perichondrium, whereas signal is absent in the chondrocytes. Black double-headed arrows delineate domains of gene expression. White arrows indicate the prehypertrophic and hypertrophic chondrocytes in Col II and Col IX samples, and the hypertrophic chondrocytes in IHH samples. White bars in WT samples define the primary ossification center that anomalously binds probe. Scale bar (black bar under the WT H&E image): 2 mm for WT samples, 1 mm for Fzd7^-C and Vangl2 samples, except 400 µm in all Col I hybridizations.

• Supplemental Figure S3 -

  Fig. S3. High-magnification images of the data from Fig. 3C. Representative regions of the developing chick growth plate were imaged from uninfected tissue (A,D,G,J) and from limbs infected with low titer (B,E,H,K) or high titer (C,F,I,L) virus. Note that injection of low titer virus results in infection primarily of the perichondrium (compare black arrows in A and B), but does not lead to significant infection of the cartilage (white arrows). By contrast, injection of high titer virus (C) results in total infection of the cartilage (white arrow). Scale bar: 120 µm in A-C; 50 µm in D-L.