DEV008599 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Expression of the CNCC markers plexin A2, FoxC1 and Crabp1 in E10.5-E12.5 control and Ltbp-1L^-/- embryos. (A-D) Tissue-slide (A,B) and whole-mount (C,D) in situ hybridization of E10.5 (A,B) and E12.5 (C,D) control and Ltbp-1L^−/− hearts, using a plexin A2 ribo-probe. Post-migratory CNCC express plexin A2 (arrows in A and C) in control but not mutant OFT. (E-H) Tissue-slide (E,F) and whole-mount (G,H) in situ hybridization of E10.5 (E,F) and E12.5 (G,H) control and Ltbp-1L^−/− hearts, using FoxC1 ribo-probe. Post-migratory CNCC express FoxC1 (arrows in E and G) in control but not Ltbp-1L^-/- OFT. (I,J) Expression of Crabp1 in E10.5 control (I) and Ltbp-1L mutant (J) embryos, assessed by whole-mount in situ hybridization. Crabp1 expression is limited only to migratory CNCC both in the control and mutant embryo. Hearts outlined in the lower-magnification images are shown at higher magnification to show lack of Crabp1 expression by the post-migratory cells in the OFT of both control and Ltbp-1L mutant embryos.

• Supplemental Figure S2 -

  Fig. S2. Tgf-β2 and Tgf-β3 distribution in the OFT of E11.5 control and Ltbp-1L^-/- embryos. (A,B) Immunofluorescence using Tgf-β2 antibody that recognizes Tgf-β2 and Tgf-β3 isoforms, shows increased amounts of Tgf-β2 and Tgf-β3 in Ltbp-1L^−/− hearts (B), especially in the mesenchymal cushions of the OFT image fields outlined by red squares and blown up to visualize post-migratory CNCC in control (A) and Ltbp-1L mutant (B) OFT.