Figure 12.

Role of IL‐6 and TNF‐α in cellular responses to TLR2 stimulation. A through C, Cellular responses to Pam3CSK4, a specific synthetic ligand for TLR2, under treatment of anti–IL‐6 antibodies (Ab) or control antibodies were assessed after 48‐hour stimulation in cardiomyocytes (A) and after 24‐hour stimulation in cardiac fibroblasts (B) and vascular endothelial cells (C). A, Cell surface areas of cardiomyocytes. Cell surface areas of 25 cardiomyocytes were measured in specimens with anti‐sarcomeric α‐actinin staining in each group. Scale bars=20 μm. B and C, Proliferation of fibroblasts (B) and vascular endothelial cells (C). Cell proliferation was assessed by means of dimethylthiazol‐carboxymethoxyphenyl‐sulfophenyl‐tetrazolium (MTS) assay and was expressed as the percentage of the absorbance to the well with cells treated by control and control antibodies. Data were obtained from 6 independent experiments in fibroblasts and 7 independent experiments in vascular endothelial cells. *P<0.05 vs control; †P<0.05 vs control antibodies. D through F, Cellular responses to Pam3CSK4, a specific synthetic ligand for TLR2, under treatment of anti–TNF‐α antibodies or control antibodies were assessed after 48‐hour stimulation in cardiomyocytes (D) and after 24‐hour stimulation in cardiac fibroblasts (E) and vascular endothelial cells (F). D, Cell surface areas of cardiomyocytes. Cell surface areas of 25 cardiomyocytes were measured in specimens with anti‐sarcomeric α‐actinin staining in each group. Scale bars=20 μm. E and F, Proliferation of fibroblasts (E) and vascular endothelial cells (F). Cell proliferation was assessed by means of MTS assay and was expressed as the percentage of the absorbance to the well with cells treated by control and control antibodies. Data were obtained from 4 independent experiments in fibroblasts and 6 independent experiments in vascular endothelial cells. *P<0.05 vs control; ^†P<0.05 vs control antibodies. IL indicates interleukin; TLR, Toll‐like receptor; TNF, tumor necrosis factor.