Figure 6.

Suspension state activated Ca²⁺/CaN/NFAT2 signaling contributing to COX-2 upregulation. (A) Cytosolic Ca²⁺ levels were estimated by Fluo-3/AM in adherent or suspension cultured MDA-MB-231 cells (scale bar = 20 μm). COX-2 protein level in suspension cultured MDA-MB-231 cells with Ca²⁺/CaN signing inhibitor CsA (B) or LaCl₃ (C). GAPDH was used as loading control. (D) Detection of NFAT1 and NFAT2 mRNA levels in adherent or suspension MDA-MB-231 cells. Values are presented as mean ± SE (n=3). *P< 0.05. (E) Detection of total protein expression and nuclear accumulation of NFAT2 in adherent or suspension MDA-MB-231 cells. (F) Detection of nuclear NFAT2 protein levels in suspension cells with CsA (1 μg/mL) or LaCl₃ (100 μM).GAPDH was used as loading control for total and cytoplasmic protein. Lamin B was used as loading control for nuclear protein. (G) Schematic of Ca²⁺/CaN/NFAT2 signaling contributing to COX-2 expression. Ad: adherent; Susp: suspension; Ctrl: control.