Crocin Suppresses LPS-Stimulated Expression of Inducible Nitric Oxide Synthase by Upregulation of Heme Oxygenase-1 via Calcium/Calmodulin-Dependent Protein Kinase 4
Figure 7
Regulation of Nrf2 activation by CAMK4. (a) RAW 264.7 cells were incubated with BAPTA/AM (B/A, 5 μM), calmidazolium chloride (CC, 5 μM), and nifedipine (Nif, 5 μM) for 1 h and then treated with crocin (500 μM) for 3 h. Equal amounts of nuclear proteins were subsequently analyzed by Western blotting. (b) Cells were transfected with pcDNA3.1 (mock, M), CAMK4 constitutive active (dCT), or kinase dead (K75E) construct and then treated with crocin (500 μM). Equal amounts of nuclear proteins were then analyzed by Western blotting. HDAC was used as a loading control. (c) Cells were cotransfected with ARE-luciferase construct and pcDNA3.1, dCT, or K75E and then treated with crocin (500 μM). Equal amounts of cell extracts were assayed for dual luciferase activity. versus the pcDNA3.1-transfected (vehicle-treated) group; versus the pcDNA3.1-transfected (crocin-treated) group.
