Nfatc1 ab2722

AS-605240 (purity>99%) was obtained from MCE (Shanghai, China). Alpha‐minimum essential medium (αMEM), trypsin-ethylene diamine tetraacetic acid (EDTA) solution, and penicillin/streptomycin (P/S) solution were purchased from Bio-Channel (Nanjing, China). We procured fetal bovine serum (FBS) from Avantor (Molendinar, Australia). Cell counting kit‐8 (CCK‐8) was supplied by MCE (Shanghai, China). We purchased mouse recombinant M‐CSF and RANKL from R&D Systems (Minneapolis, USA). We purchased primary antibodies against the total and phosphorylated proteins, including β‐actin (#4970, 1:1000), PI3K (#4295, 1:1000), p-PI3K (#17366, 1:1000), Akt (#4691, 1:1000), p-Akt (#4060, 1:1000), P65 (#8242, 1:1000), p-P65 (#3033, 1:1000), IκBα (#4814, 1:1000), p-IκBα (#2859, 1:1000), P38 (#8690, 1:1000), p-P38 (#4511S, 1:1000), ERK (#4695, 1:1000), p-ERK (#4370, 1:1000), JNK (#9252, 1:1000), p-JNK (#9255, 1:1000) and RUNX2 (#12556, 1:1000) from Cell Signaling Technology (Danvers, USA). We obtained c-Fos (ab222699, 1:1000) and NFATc1 (ab2722, 1:1000) from Abcam (Shanghai, China), and the dilution of the primary antibodies was obtained from Beyotime Biotechnology (Shanghai, China). We purchased a secondary antibody (#S0001, 1:20000) from Affbiotech (Changzhou, China).

SSD with >98% purity was obtained from MedChemExpress (NJ, USA). Dimethyl sulfoxide (DMSO) (Meilunbio, Dalian, China) was used to dissolve SSD at a concentration of 1 mmol/L and was stored at −20°C. The culture medium was used for further dilutions in the in vitro experiments, and phosphate-buffered saline (PBS) was used for in vivo experiments. Alpha-modified Eagle medium (α-MEM) was provided by HyClone Laboratories (Logan, UT, USA). Foetal bovine serum (FBS) was obtained from Gibco-BRL (Gaithersburg, USA). Cell counting kit-8 (CCK-8) was obtained from Beyotime (Shanghai, China). M‐CSF (mouse recombinant) and RANKL were provided by R&D Systems (Minneapolis, USA). Primary antibodies, including β‐actin (#4970), P65 (#8242), p-P65 (#3033), P38 (#8690), p-P38 (#4511S), AKT (#4691), p-AKT(#4060), ERK(#4695), p-ERK(#4370), IκBα(#4814), p-IκBα (#2859), PI3K(#4295), p-PI3K (#17366), JNK (#9252) and p-JNK(#9255), were provided by Cell Signaling Technology (1:1000, Danvers, USA). In addition, c-Fos(ab222699) and NFATc1(ab2722) were obtained from Abcam (1:1000, Cambridge, UK), and the dilution of the primary antibodies was purchased from Beyotime Biotechnology.