Fig. 5.

Nell-1 inactivation reduces CNCCs osteogenic capacity and cell proliferation. The CNCCs isolated from Nell-1^Wnt1KO newborn mice demonstrated significantly impaired osteogenic capacity as demonstrated by a ALP staining and mineralization, and b gene expression of the osteogenic markers ALP, OCN, and Runx2; c in vitro EdU labeling exhibited significantly reduced positive cells from Nell-1^Wnt1KO mice, and d flow cytometry cell cycle analyses further confirmed the lower proliferative cell populations in Nell-1^Wnt1KO mice. Specifically, CNCCs in the DNA synthesis phase (S phase) of WT mice (11.8 ± 0.27%) were significantly more than that of Nell-1^Wnt1KO (7.2 ± 0.66%). e While the rates of early apoptosis of CNCCs measured by flow cytometry from Nell-1^Wnt1KO mice (9.0 ± 0.13%) was slightly higher than that of WT mice (8.5 ± 0.20%), no statistically significant difference was found. NS no significance; *P < 0.05; ***P < 0.005; ****P < 0.001