Xenbase Image ID: 139653

Figure 4. Sox5 Recruits Smad1/4 to Target Promoters through Physical Association (A) GST pull-down assay using lysates from uninjected embryos or embryos injected with Sox5 or Sox6 mRNA assaying for binding to GST, Smad1-GST, Smad3-GST, or Smad4-GST. Western blots show expressed levels of Sox5 and Sox6. (B) GST pull-down assay using lysates from embryos injected with Sox5 mRNA assaying for binding to GST, Smad1-GST, Smad1-MH1-GST, or Smad1-MH2-GST. (C) Schematic representations of Smad1, Smad1-MH1, and Smad1-MH2 constructs used in pull-down assays. (D) Coomassie staining of expressed GST fusion proteins used for pull-downs in (A) and (B). (E) ChIP-qPCR analysis of Smad1 enrichment on EPK promoter relative to eEF1α promoter. Smad1 enrichment is significantly reduced when Sox5 is depleted. Error bars represent SEM of three independent biological experiments. p = 0.0022. (F) ChIP q-PCR analysis of Smad1 enrichment on Vent2 promoter relative to control eEF1α promoter. Smad1 enrichment is significantly reduced when Sox5 is depleted. Error bars represent SEM of three independent biological experiments. p = 0.0472. (G) ChIP analysis examining Sox5 enrichment on EPK promoter relative to eEF1α promoter. Smad1 enrichment is significantly reduced in the presence of Chordin. Error bars represent SEM of three independent biological experiments. p = 0.0048. (H) ChIP analysis examining Sox5 enrichment on Vent2 promoter relative to eEF1α promoter. Sox5 enrichment is significantly reduced in the presence of Chordin. Error bars represent SEM of three biological experiments. p = 0.0006. See also Figure S4. Image published in: Nordin K and LaBonne C (2014) Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

Return to previous page