Plcγ2

Methyl gallate (98% purity) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in dimethyl sulfoxide (DMSO). Recombinant soluble human macrophage colony-stimulating factor (M-CSF) and RANKL were obtained from PeproTech EC Ltd. (London, UK). A monoclonal antibody against β-actin was obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against phosphorylated (phospho)-p38, p38, phospho-extracellular signal-regulated protein kinase (ERK) 1/2, ERK 1/2, phospho-JNK, JNK, phospho-IκB, phospho-Akt, Akt, and Bruton’s tyrosine kinase (Btk) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Antibodies against c-Fos, nuclear factor of activated T cells c1 (NFATc1), IκB, phospho-PLCγ2, and PLCγ2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). An antibody against phospho-Btk was purchased from GeneTex (Irvine, CA, USA). Two kinds of secondary antibodies including horseradish peroxidase–conjugated sheep anti-mouse IgG antibody and donkey anti-rabbit immunoglobulin antibody were obtained from Enzo Life Sciences (Farmingdale, NY, USA). Fetal bovine serum (FBS), α-minimum essential medium (α-MEM), penicillin, and streptomycin were purchased from Gibco BRL (Grand Island, NY, USA). All other chemicals were of analytical grade or complied with the standards required for cell culture experiments.