Fig. S7

Knockdown of Fscn1a impairs sqt ectopic expression-induced exression of target genes.

(a-b) mammalian Fscn1 is required for Activin signal transduction. NIH3T3 (a) and NMuMG (b) cells transfected with the indicated plasmids (2 µg shRNA plasmids together with or without 0.5 µg reporter plasmids in one well of 6-well plate) were treated with Activin A (25 ng/ml) for 12 hours before harvested for luciferase measurements. The relative luciferase activity was the mean with SD from three independent experiments. Student’s t test, *P < 0.05, **P < 0.01. (c-e) fscn1a is essential for Nodal signal to regulate target gene expression. Wild-type embryos were injected with indicated MOs and mRNAs and harvested at shield stage for in situ hybridization (c and d) and real-time PCR experiments (e). Note that the sqt overexpression induced- but not casmad2 overexpression induced-expression of Nodal signal target genes was inhibited by fscn1a MO1 injection. Scale bar, 200 µm. In panel e, the expression of β-actin was used as a reference to normalize the amount of mRNAs in each sample. The data are presented as mean ±SD of three independent experiments. Student’s t test, *P < 0.05, **P < 0.01; NS, nonsignificant. UIC, uninjected control.