Extended Data Figure 5. MMP2 and 14 expression is regulated by Nodal.

A) Expression levels for extracellular matrix (ECM) genes are plotted on the UMAP for all cell types from E5.25-E6.5(n=1724 cells). All expression values are log(counts). B) Representative examples of fluorescent RNA in-situ for control (n=3) and SB431542-treated embryos(n=3). Embryos were recovered at E5.75 and cultured for 18 hours before fixation. Nodal signalling is inhibited after treatment with SB431542 evident from the absence of Nodal and Tdgf1 mRNA. Scale bars = 20um. C) Representative examples of control(n=5) and Nodal inhibitor-treated(n=5) (18 hours) embryos stained for MMP14. D) Western blot analysis for MMP2 and MMP14 in control and Nodal-inhibited (SB431542 treated) ESCs. n=3 independent experiments. For gel source data see Supplementary Figure 1. E) Quantification of relative protein levels based on western blots of ESCs in the presence or absence of Nodal inhibitor SB431542. Two sided unpaired student’s t-test. MMP2: **p=0.0088; MMP14: **p=0.0086; n=3 independent experiments; mean±SEM. F) ChIP tracks for Smad2/3. Smad2/3 binding on MMP2 & 14 is lost upon Nodal inhibition (green rectangles). Scale bars=20um