Fig. 3
Co-incubation of EMPA (EMPA) and doxorubicin (DOXO) reduced the expression of pro-inflammatory interleukins, leukotrienes and p65/NF-Kb compared to only DOXO-treated cells. For each experiment, 5 × 103 cells/well were seeded in a 96-well plate; cells were exposed to EMPA (10, 50, 500 nM), LPS (40 ng/ml), DOXO (100 nM) alone or with EMPA at 10, 50 and 500 nM (n = 4/group). Expression of (A) Interleukin-8, (B) Interleukin-6 and (C) Interleukin1-β (pg/mL). D Leukotriene B4 expression (pg/mL) in cardiomyocytes exposed to arachidonic acid (10 µM), EMPA (10, 50, 500 nM) alone or associated to arachidonic acid, DOXO (100 nM) alone or co-incubated with EMPA at 10, 50 and 500 nM. E p65/NF-kB DNA binding activity, expressed as fold of untreated (control) cells (n = 3). (F) MyD88 and NLRP3 expression (fold of control), (n = 3); One-way ANOVA. Value are expressed ± SD. ***P < 0.001; **P < 0.01; *P < 0.05; ns: not significant. G: Confocal laser scanning microscope images of cardiomyocytes exposed to medium alone (A), EMPA 100 nM (B), DOXO 100 nM (C) and EMPA (100 nM) DOXO (100 nM) in combination (D) for 24 h (n = 3/group). Green fluorescence indicates p65/NF-kB staining; red fluorescence indicates cell nucleus. Scale bar: 50 µM