HIF1 regulation of Sox9 is necessary to maintain differentiation of hypoxic prechondrogenic cells during early skeletogenesis

DEV008441 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 - Fig. S1. Aberrant joint formation in Prx1-Hif1α limbs. (A,B) H&E staining of E13.5 control (A) and Prx1-Hif1α (B) limbs. H, humerus; R, radius; U, ulna. Cell death is evident in E13.5 Prx1-Hif1α elbow joint. (C-F) Higher magnification of articular cartilage region of Prx1-Hif1α (D) demonstrates loss of the typical morphology observed in the control (C); fused joints are observed in E18.5 Prx1-Hif1α digits (F) compared with control (E). (G-I) Tunnel assay demonstrates massive cell death in E15.5 Prx1-Hif1α elbow (H) relative to control (G); only minor cell death is detectable in E15.5Prx1-Hif1α digits (I). (J,K) Histology sections of E15.5 autopod of control (J) and Prx1-Hif1α (K) were stained with Alcian Blue to detect chondrocyte differentiation. Arrows indicate undifferentiated chondrocytes that did not stain with Alcian Blue.

• Supplemental Figure S2 -

  Fig. S2. Chromatin immunoprecipitation assay of the mouse Pgk1 gene. Sheared chromatin from hypoxic (1% oxygen) or normoxic (20% oxygen) micromass culture cells was precipitated with polyclonal anti-HIF1α or control IgG. Binding was indicated by the percentage of total input chromatin DNA (n=2).