Fig. 5. TRPV1-mediated Ca²⁺ influx promotes Nrf2 nuclear translocation in RAW264.7 cells.

A Fluorescence images and B their quantitative analysis of intensity in the Fluo-4 AM-loaded RAW264.7 cells treated with CaCl₂, CPS, or CaCl₂ + CPS under the Ca²⁺-free medium for 10, 30, 60, and 90 min. Scale bars: 50 μm. C Western blot analysis of cytosolic and nuclear Nrf2 protein levels in the cells treated with LPS, LPS + CPS, and LPS + CPS with EDTA (left) or Ca²⁺-free medium (right) for 3 h. D Quantitative analysis and E representative images of Nrf2 immunofluorescence in the cells treated with LPS, LPS + CPS, and LPS + CPS with EDTA or Ca²⁺-free medium for 3 h. Scale bars: 10 μm. White arrow: cells with Nrf2 nuclear translocation. Data (n = 3) are shown as mean ± SD. *P < 0.05; ***P < 0.001; NS nonsignificant.