Figure 3.

NFATc1 Is a Rapidly Degraded Nuclear Protein

A, Western analysis of NFATc1 expression. RAW264.7 cells were treated with sRANKL (300 ng/ml) for 24 h and then subjected to an additional treatment with CHX (10 μg/ml). Cells were isolated at the times indicated and fractionated to obtain cytosolic and nuclear components, and the components were then subjected to Western blot analysis using antibodies to NFATc1. The arrow indicates the presence of NFATc1 protein at approximately 90 kDa. B, Determination of NFATc1 half-life. The signals generated in panel A were quantitated densitometrically and normalized to the levels present before CHX administration. NFATc1, t_1/2 = 3.1 h; r² = 0.97. These data are representative of three independent experiments.