Fig. 5

c-FOS mRNA and protein expression are dependent on MEF2C in human and mouse OCPs. Human osteoclast precursor cells were nucleofected with control or MEF2C #2 siRNAs. a RT-qPCR analysis of human FOS mRNA after 24 h of culture with or without RANKL normalized relative to TBP mRNA (control samples without RANKL set at 1.0). n = 5. b Representative immunoblotting of c-FOS in nuclear lysates. Lamin B1 and α-tubulin were used as controls for nuclear and cytoplasmic proteins, respectively. c Densitometric quantitation of c-FOS band intensity after 24 h of culture with RANKL from three independent donors. Mouse osteoclast precursor cells from MEF2C^ΔMX KO or littermate control WT mice were cultured with M-CSF and RANKL. d RT-qPCR analysis of mouse Fos mRNA after 24 h of culture with or without RANKL (50 ng·mL⁻¹) normalized relative to Hprt mRNA (control samples without RANKL set at 1.0). WT; n = 11, KO; n = 12. e Representative images of immunoblotting analysis of c-FOS expression in nuclear lysates. Right panel, densitometric quantitation of band intensity from four samples of each genotype. f Densitometric quantitation of c-FOS band intensity (n = 4). Human osteoclast precursor cells were transduced with adenoviral particles encoding GFP or MEF2C-FLAG and stimulated with RANKL. g RT-qPCR analysis of human Fos mRNA after 12 h of culture with or without RANKL (40 ng·mL⁻¹) normalized relative to TBP mRNA (control samples without RANKL set at 1.0). n = 8. h Immunoblotting of c-FOS at the indicated times. Representative images from five independent experiments. i Densitometric quantitation of c-FOS band intensity from five independent experiments. Data are shown as mean ± SD. Statistics used: a, d, g, i repeated measurement two-way ANOVA, c, f paired t-test. *P < 0.05, **P < 0.01