Fig. 2

PA leads to unstability of mitochondria member potential triggered cell apoptosis through mitochondria dependent pathway and NFκB signaling pathway. a H9c2 cells were treated with PA (0.5 mM) for 24 h. △Ψm was assessed with signal from monomeric and J-aggregate JC-1 fluorescence. JC-1 fluorescence was measured by flow cytometry. Left: control, Right: PA. b H9c2 cells were treated with PA (0.5 mM) for indicated time. p-Akt, Bcl-2, Bax, caspase 3 expression was estimated by immunoblotting.(c) H9c2 cells were treated with PA (0.5 mM) for 24 h and the cell lysates were fractionated into cytosolic and mitochondrial proteins. Cytochrome c was analyzed by immunoblotting. β-actin and COX IV served as the cytosolic and mitochondrial loading controls. d H9c2 cells were treated with PA (0.5 mM) for indicated time. The expression of MAPK family (p-ERK, p-JNK, p-P38) was analyzed by immunoblotting. e H9c2 cells were incubated with PA (0.5 mM) for 0-2 h. The expression of NFκB, and IκB was analyzed by immunoblotting. β-actin and PCNA served as the cytosolic and nuclear loading controls. f Cells were transfected with a luciferase NFκB reporter construct. After transfection and treatment with PA for indicated time (0, 0.5, 1 or 2 h), the cells were assayed for luciferase activity. *p < 0.05 compared with the control. g After H9c2 cells were transfected with JNK1, NFκB siRNA (10 nM) for 24 h, followed by treatment of PA for 24 h, scramble for nonspecific siRNA control. The levels of proteins indicated were analyzed by Western blot