Fig. 7

Coculture with the preosteoblastic cell line MC3T3 rescues impaired osteoclastogenesis induced by Fanca KO. A Number of OCs per field from WT and Fanca^−/− cells plated on WT or Fanca-depleted MC3T3 cells at day 4 of differentiation. B Relative Nfatc1, Cathepsin K and Cdkn1a expression evaluated by qRT-PCR in OCs from WT and Fanca^−/− mice cocultured with WT or Fanca-depleted MC3T3 cells at day 4 of differentiation. C Western blot showing Fanca, Nfatc1 and Cathepsin K in OCs from WT and Fanca^−/− mice at day 4 of differentiation plated on WT or Fanca-depleted MC3T3 cells. β-Actin was used as a loading control. Middle and right: Relative protein expression of Nfatc1 and Cathepsin K in OCs from WT and Fanca^−/− mice at day 4 of differentiation plated on WT or Fanca-depleted MC3T3 cells. D Relative TNFα level in the WT or siFanca-depleted MC3T3 cells as evaluated by FACS analysis. E Number of OCs per field from Fanca^−/− cells plated on WT or Fanca-depleted MC3T3 cells and left untreated (NT) or treated with an anti-TNFα neutralizing antibody (10 ng/ml) at day 4 of differentiation. Data are shown as the mean ± SEM. Each small circle represents an individual analyzed mouse or individual experiment (D). Statistical analysis was performed by t tests: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001