XB-IMG-159710
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Fig. S2: RNA-seq of DMSO and DMH1 treated embryos identified BMP-regulated genes.
(A) Scatter plot showing log2FC in expression between DMSO and DMH1 treated FG (left) and between
DMSO and DMH1 treated HG (right) samples. Transcripts are colored based on the normal control
expression; HG-enriched in green, FG-enriched in orange, genes expressed similarly in FG and HG
(FG=HG) in grey and normally not expressed in control FG or HG in black. (B) Venn diagram illustrates
overlap between transcripts up regulated (é) or down regulated (ê) upon DMH1 treatment in FG and HG
tissues. FG or HG transcripts repressed upon DMH1 treatment (log2FC ≤-1, FDR ≤5% relative to
controls) are considered to be BMP-activated genes (n=697), whereas FG or HG transcripts that are
increased upon DMH1 (log2FC ≥1, FDR ≤5%) are classified as BMP-repressed genes (n=1063). Eight
transcripts had ambiguous regulation being both activated and repressed by DMH1 in FG or HG tissues,
and were excluded from further analysis. Overall we categorized a total of 1760 (697+1063) BMPregulated
genes in the FG and HG tissue. (C) GO term analysis of BMP-activated and -repressed genes.
(D) Unsupervised clustering of BMP-regulated genes in control (CONT) and DMH1 treated FG and HG samples compared to dorsal explants (DORS), which contain a thin layer of dorsal endoderm (yellow)
along with neural and somite tissue. The DMH1-treated FG showed similarities to the dorsal tissue
suggesting that BMP induces ventral mesendoderm fate and represses dorsal fate. (E) In situ
hybridization of control, DMH1 treated or BMP2 injected embryos, in a cross section confirms that
expression of the dorsal mesoderm gene foxc2 is expanded ventrally with DMH1 and restricted dorsally
upon BMP injection. White line indicates expression domain. Image published in: Stevens ML et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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