Fig. 6

Bhlhe40 deficiency leads to a decrease in c-Fos/Nfatc1 and directly targets Fos and Nfatc1. A The correlation expression of Bhlhe40 and Nfatc1 (GSE51495 dataset) (n = 30). B The correlation expression of Bhlhe40 and Fos (GSE7158 dataset) (n = 26). C, D Immunofluorescence and quantitative (n = 5) of NFATc1 (green) in BMMs or OCs between Wt and Bhlhe40^−/− mice. BMMs were stained with CD11b (red). OCs were stained with CTSK (red). Nucleus was stained with DAPI (blue) (Scale bar, 150 μm). E, F Immunofluorescence and quantitative (n = 5) of c-Fos (green) in BMMs or OCs between Wt and Bhlhe40^−/− mice. BMMs were stained with CD11b (red). OCs were stained with CTSK (red). Nucleus was stained with DAPI (blue) (Scale bar, 150 μm). G, H Immunohistochemistry of NFATc1 and quantitative analysis (n = 5) in femurs from Wt and Bhlhe40^−/− mice (Scale bar, 50 μm). I, J Immunohistochemistry of c-Fos and quantitative analysis (n = 5) in femurs from Wt and Bhlhe40^−/− mice (Scale bar, 50 μm). K Immunohistochemistry of NFATc1 in femurs from each group of mice for bone marrow transplantation (Scale bar, 50 μm). L Immunohistochemistry of c-Fos in femurs from each group of mice for bone marrow transplantation (Scale bar, 50 μm). M Anti-BHLHE40 ChIP assay for the Nfatc1 promoter in BMMs. Predicted anti-BHLHE40 binding sites are indicated. Immunoprecipitated DNA was amplified by qPCR (n = 3). Normal rabbit IgG was used as a negative control. Results are presented as ChIP/Input. N Anti-BHLHE40 ChIP assay for the Fos promoter. Predicted Anti-BHLHE40 binding sites are indicated. Immunoprecipitated DNA was amplified by qPCR (n = 3). Normal rabbit IgG was used as a negative control. Results are presented as ChIP/Input. All data are mean ± SD; ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001. by unpaired Student’s t test, two-tailed and one-way ANOVA followed by Tukey’s post hoc test