Fig 2: Calcineurin/Akt/mTOR/NFATC2 signalling controlled by store-operated Ca2+ entry. (A) M-CSF/RANKL-induced calcineurin activity in BMDMs in the presence or absence of CsA. (B-G) Phosphorylation levels of (B) mTORS2448, (C) AktT308, (D) AktS473, (E) p70S6S235/236 and dephosphorylation levels of (F) NFATC2 and (G) total NFATC2 were detected by flow cytometry prior to and after treatment of STIM1wt or STIM1mut BMDMs with M-CSF/RANKL. (H) The CSFE fluorescence intensity, as well as the expression of ki67, cathepsin K and MMP9 in STIM1wt or STIM1mut BMDMs were detected prior to and after treatment with M-CSF/RANKL in the presence or absence of 7 nM CsA, 0.1 nM Rap or NFATC2 siRNA. (I) Proliferation and (J) differentiation of STIM1wt or STIM1mut BMDMs detected prior to and after treatment with M-CSF/RANKL in the presence or absence of 7 nM CsA, 0.1 nM Rap or NFATC2 siRNA. Values are expressed as the mean ± standard deviation (n=6). **P<0.01 and ***P<0.001 vs. Untreated BMDMs. BMDMs, bone marrow-derived mononuclear macrophages; CSFE, carboxyfluorescein succinimidyl ester; STIM1, stromal interaction molecule 1; wt, wild-type; mut, mutant; MMP, matrix metallopeptidase; TRAP, tartrate-resistant acid phosphatase; RANKL, receptor activator of NF-?B ligand; M-CSF, macrophage colony-stimulating factor; siRNA, small interfering RNA; CsA, cyclosporin A; Rap, rapamycin; p-Akt, phosphorylated Akt.