Enhanced chondrogenesis and osteogenesis in Novdel3-/- PEFs. A-H: Staining of micromass cultures of PEFs derived from wild type (A-D) and Novdel3-/- (E-H) embryos. Alcian blue staining after 5 days (A, E) and 9 days (B, F) in culture showing enhanced cartilage differentiation in Novdel3-/- cells, with the dashed line in B and F marking the extent of the micromass. Alkaline phosphatase staining after 5 days (C, G) and 9 days (D, H) in culture showing enhanced osteoblast differentiation in Novdel3-/- cells. I-L: Morphology of monolayer PEF cultures derived from wild type (I, J) and Novdel3-/- (K,L) embryos with (I, K) and without (J, L) phase contrast. Alkaline phosphatase staining after 10 days in culture (I, J, K, L) showing osteoblast differentiation in the Novdel3-/- PEFs but not in the wild type PEFs. Scale bar in I-N = 10 μm. M: Semi quantitative RT-PCR of mRNA from wild type (+/+) and Novdel3-/- PEFs using primers for alkaline phosphatase, collagen I and osteocalcin. Two fold serial dilutions of cDNA were used and normalised to gapdh.