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. 2014 Nov 15;141(22):4285–4297. doi: 10.1242/dev.110908

Fig. 5.

Fig. 5.

The EMT master regulator Snai1 is a direct target of Msgn1. (A) Semi-quantitative PCR analysis of Snai1 expression in iF-Msgn1 EBs treated with (+) or without (−) Dox from 0-72 h. Cyclophilin A (PpiA) is used as a loading control. (B) Msgn1 binds to the +5148 bp enhancer of Snai1. The schematic (below) shows the two E-boxes (red rectangles) identified in the Snai1 +5148 enhancer peak. Arrowheads indicate orientation and y-axis denotes peak height, corresponding to the normalized fold enrichment. (C) Luciferase reporter assay of Snai1 +5148 enhancer co-transfected with control or Msgn1 expression constructs. The y-axis shows the normalized fold change; error bars show the s.d.; n=3. (D) ChIP-qPCR analysis of Msgn1 enrichment on Snai1 +5148 locus using control ‘C’ and anti-Msgn1 ‘αM’ ascites in E9.5 PSM and head extracts and in iF-Msgn1 EBs. (E-H) WISH analysis of Snai1 mRNA in E8.5 control and Msgn1−/− embryos (E,G, lateral view; F,H, ventral view). Scale bars: 200 μm.