Cell organization, growth, and neural and cardiac development require αII-spectrin

JCS080374 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Whole embryo expression of αII-spectrin. Whole mount of an E14 heterozygous embryo stained for expression of wild-type αII-spectrin or for the αII-spectrin/β-gal fusion.

• Supplemental Figure S2 -

  Fig. S2. Spectrin-binding proteins unchanged in Spna2^−/−embryos. Results from two pairs of animals shown. The levels of ena/VASP, Abi, Kap3, and 14-3-3 are all unchanged by the disruption of αII-spectrin. These proteins all have been shown to bind directly to αIIβII-spectrin.

• Supplemental Figure S3 -

  Fig. S3. The level of Smad proteins are unchanged in Spna2^−/−embryos and MEFs. (A) The overall levels of the indicated Smad proteins and phosphoSmad2 (pSmad2) in E13.5 embryos were examined by western blotting. Three pairs of normal and αII-spectrin-deficient animals were examined. Relative to actin, no significant changes were detected. (B) The response of 3T3 fibroblasts and cultured MEF's from Spna2^+/+ or Spna2^−/− embryos to TGF-β stimulation (0 to 10±ng/ml). All cells responded to TGF-β with increased pSmad2. No differences were detected between wild-type and the αII-spectrin-deficient cells.

• Supplemental Figure S4 -

  Fig. S4. Vimentin staining is enhanced in the SVZ of spectrin deficient mice. Sections of brain from E13.5 embryos were stained by immunoperoxidase with antibody to vimentin. Note the enhanced density and intensity in the spectrin-deficient brain (Spna2^−/−). Vimentin is a marker of glial differentiation.

• Supplemental Table S1 - Primers used for Q-RTPCR.