Figure 5.

Cellular calcium is involved in Ang ІІ-induced inflammation. (A) HL-1 cells were pre-treated with fluo-3 AM for 30 min and then with 1 μM Ang ІІ. Green fluorescence was detected using a confocal microscope. (B) HL-1 cells were treated with Ang ІІ (1 μM) for 24 h. Phosphorylated CaMKІІ and CaMKІІ protein levels were assessed by western blot analysis with a specific antibody. (C) Cells were pre-treated with KN-93 (5 μM) for 30 min and treated with Ang ІІ for 24 h. Cell lysates were analysed by western blotting analysis with antibodies against phosphorylated JNK. Non-phosphorylated JNK was used as a control. β-actin served as a protein loading control. (D) Cells were transiently transfected with CaMKІІδ siRNA (100 nM) for 2 days and then treated with Ang ІІ (1 μM) for 24 h. CaMKІІδ protein levels were assessed by western blot analysis with a specific antibody. (E) HL-1 cells were transiently transfected with CaMKІІδ siRNA and then the cells were lysed, and the expression of p-JNK, JNK, and β-actin was quantified by western blot analysis. Levels of JNK and β-actin were also measured as protein loading controls. (F) Cells were treated with KN-93 and then incubated with Ang ІІ. The protein expression of TGF-β1, NF-κB, and β-actin was evaluated by western blot analysis. Blotting with an antibody specific to β-actin served as a control. (G) CaMKІІδ siRNA was transiently transfected into the cells, followed by Ang ІІ treatment. The cell lysates were analysed by western blot analysis with TGF-β1, NF-κB, and β-actin antibodies. *P < 0.05, **P < 0.01, compared with the untreated controls. Results are from three independent experiments. Blots are displayed in cropped format.