Figure 3. BMP Signaling Requires Sox5 to Activate Transcriptional Targets (A and B) Ectoderm explant assay examining Vent1 (A) and Msx1 (B) in explants injected with BMP4 mRNA or BMP4 mRNA with Sox5 MO. Explants were collected alongside sibling embryos cultured until early neurula stages (stage 14). (C) Western blot using lysates from embryos injected either with BMP4, BMP4/Sox5, BMP4/Sox5MO, or Chordin to examine levels of phosphorylated Smad1 and unphosphorylated Smad1. (D) Western blot using using lysates from embryos injected with Activin, Activin/Sox5, Activin/Sox5MO, Sox5, or Sox5MO to examine levels of phosphorylated Smad2. (E and F) Luciferase assay examining fold activation by the Vent2 promoter, p = 0.0068 (E), or 12XGCCG reporter, p = 0.0227 (F), in the presence of active Smad1/4 complexes or with active Smad1/4 and Sox5 MO. (G) Luciferase assay using the Vent2 reporter and the Vent2 3x Mutant reporter (carrying the same mutations as in Figure S3C). The mutations that reduce Sox5 binding also reduce activation of the reporter by active Smad1/4. Error bars represent SD of three independent experiments. See also Figure S3.
Image published in: Nordin K and LaBonne C (2014)
Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V.
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