nfat inhibitor  (MedChemExpress)

Figure Legend Snippet: BHLHE41 directly targeted NFATc1 (A) The correlation expression of BHLHE41 and NFATc1 ( GSE56815 dataset) (n = 80). (B and C) Western blotting and quantification (n = 3) to detect the expression of NFATc1 in si-NC and si-BHLHE41 in BMMs. (D and E) Western blotting and quantification (n = 3) to detect the expression of NFATc1 in Control and LV-BHLHE41 in BMMs. (F) Motif of BHLHE41 promoter binding site. (G) Anti-BHLHE41 ChIP assay for the NFATc1 promoter in BMMs. Predicted Anti-BHLHE41 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. (H and I) TRAP staining and quantification (n = 3) to detect the osteoclast differentiation of BMMs in Control, BHLHE41 Overexpression and BHLHE41 Overexpression treated with si-NFATc1 (Scale bar, 50 μm). (J) OC-specific genes expression of BMMs in Control, BHLHE41 Overexpression and BHLHE41 Overexpression treated with si-NFATc1 (n = 3). (K and L) TRAP staining and quantification (n = 3) to detect the osteoclast differentiation of BMMs in Control, BHLHE41 Overexpression and BHLHE41 Overexpression treated with NFAT Inhibitor (Scale bar, 50 μm). (M) OC-specific genes expression of BMMs in Control, BHLHE41 Overexpression and BHLHE41 Overexpression treated with NFAT Inhibitor (n = 3). NFAT Inhibitor reduced the expression of NFATc1. All data are mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Techniques Used: Expressing, Western Blot, Control, Binding Assay, Immunoprecipitation, Amplification, Negative Control, Staining, Over Expression

Figure Legend Snippet:

Techniques Used: Virus, Over Expression, Plasmid Preparation, Recombinant, Immunoprecipitation, Protease Inhibitor, Polymer, Reverse Transcription, SYBR Green Assay, RNA Extraction, Transfection, Purification, Isolation, Software, Micro-CT, Imaging, Real-time Polymerase Chain Reaction, Inverted Microscopy, Membrane, Magnetic Beads