JCS034736 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. FGF1-induced ERK phosphorylation is affected in lung mesenchymal cells or MEFs derived from Ndst1^−/− mice. (A-D) In situ HS binding assays for lung sections from mice at 18.5 d.p.c. reveal that binding ability of FGF1 to HS is decreased in Ndst1^−/− mice (B) compared with that of normal littermates (A). (C,D) FGF1 binding following pretreatment with heparitinase. Scale bar: 100 µm. (E) Lung mesenchymal cells or MEFs were stimulated for 10 minutes at 37°C with 5 ng/ml or 50 ng/ml FGF1. ERK1/2 phosphorylation was barely detected without treatment of FGF1 in either wild-type or Ndst1 mutant cells. In response to 5 ng/ml FGF1, ERK1/2 phosphorylation was reduced in Ndst1^−/− cells. However, in response to 50 ng/ml FGF1, Ndst1^−/− cells showed increased ERK1/2 phosphorylation. Expression of ERK1/2 serves as loading control.

• Supplemental Figure S2 -

  Fig. S2. Hedgehog signaling is not affected in Ndst1 mutant lungs. Immunohistochemical staining of Patched (A,B) or Gli1 (C,D), in lungs of 18.5 d.p.c. wild-type (A,C) and Ndst1^−/− mice (B, D). (E,F) Control sections were stained with block serum instead of antibodies. Scale bar: 100 µm.