Figure 5.

Cyanidin modulates Crz1 activation in yeast and NFAT and NF-kB in microglia. (a) BY4741 yeast strain encoding CRZ1^promoter–lacZ was subjected or not to 50 µM of the indicated compounds, induced with 1.8 mM MnCl₂, and Crz1 activation was assessed by monitoring β-galactosidase activity using 2-Nitrophenyl β-d-galactopyranoside (ONPG). The immunosuppressant FK506 was used as a positive control. (b) NFATc1 and (c) NF-kB translocation to the nucleus in N9 cells pre-treated with 5 µM cyanidin or cyanidin-3-O-glucoside for 6 h before 1 h ATP stimulation (NFATc1) or LPS stimulation (NF-kB). NFATc1 and NF-kB were immunostained with Alexa Fluor 594 (red) and nuclei were stained with DAPI (blue). (d) NO and TNF-α release in N9 cells pre-treated with 5 µM cyanidin or cyanidin-3-O-glucoside for 6 h before LPS stimulation (NF-kB). Representative images are shown, and the values represent the mean ± SEM of at least three biological replicates, ** p < 0.01, *** p < 0.001.