Journal: Biomedicines

Article Title: CCL4 Deletion Accelerates Wound Healing by Improving Endothelial Cell Functions in Diabetes Mellitus.

doi: 10.3390/biomedicines10081963

Figure Lengend Snippet: Figure 1. CCL4 inhibition by siRNA recovered functions of high-glucose-impaired HDMECs with increased angiogenic protein expressions. Western blotting and statistical analyses of CCL4 ((A); n = 3), p-AKT, p-eNOS, VEGF, and SDF-1α. The angiogenic proteins were reversed after the administration of siCCL4 ((B); n = 3). Tube formation ability and quantitative analysis of HDMECs; the images were captured using a (×40) microscope. The tube formation ability was improved after the administration of siCCL4 ((C); n = 3). Migration ability and quantitative analysis of HDMECs; the images were captured using a (×100) microscope. The migration ability was improved after the administration of siCCL4 ((D); n = 3). HG represents high glucose. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were transfected with CCL4 siRNA (Santa Cruz, sc-43932, Dallas, TX, USA) using Oligofectamine (Invitrogen, 12252011, Carlsbad, CA, USA) in opti-MEM.

Techniques: Inhibition, Western Blot, Microscopy, Migration

Journal: Biomedicines

Article Title: CCL4 Deletion Accelerates Wound Healing by Improving Endothelial Cell Functions in Diabetes Mellitus.

doi: 10.3390/biomedicines10081963

Figure Lengend Snippet: Figure 2. CCL4 inhibition by siRNA recovered functions of EPCs from type 2 DM patients with increased angiogenic protein expressions. Western blotting and statistical analyses of CCL4 ((A); n = 3), p-AKT, p-eNOS, VEGF, and SDF-1α. The angiogenic proteins were reversed after the administration of siCCL4 ((B); n = 3). Tube formation ability and quantitative analysis of EPCs from type 2 DM patients and control subjects; the images were captured using a (×40) microscope. The tube formation ability was improved after the administration of siCCL4 ((C); n = 3). Migration ability and quantitative analysis of EPCs from type 2 DM patients and control subjects; the images were captured using a (×100) microscope. The migration ability was improved after the administration of siCCL4 ((D); n = 3). * p < 0.05, ** p < 0.01.

Article Snippet: Cells were transfected with CCL4 siRNA (Santa Cruz, sc-43932, Dallas, TX, USA) using Oligofectamine (Invitrogen, 12252011, Carlsbad, CA, USA) in opti-MEM.

Techniques: Inhibition, Western Blot, Control, Microscopy, Migration

Journal: Biomedicines

Article Title: CCL4 Deletion Accelerates Wound Healing by Improving Endothelial Cell Functions in Diabetes Mellitus.

doi: 10.3390/biomedicines10081963

Figure Lengend Snippet: Figure 3. CCL4 inhibition by knockout improved wound repair in diabetic mice. Representative wound areas (A). The closure rates of 3-millimeter punch biopsies were measured ((B); n = 6). Representative images with H&E staining (C). Representative images with immunostaining of CD31 and Ki67. Both CD31- and Ki67-positive areas were enhanced in the CCL4-knockout mice (D,E). Western blotting and statistical analyses of CCL4, p-AKT, p-eNOS, VEGF, and SDF-1α in the wound area. The angiogenic proteins were enhanced in tissues from the CCL4-knockout mice ((F); n = 3). Western blotting and statistical analyses of TNF-α and IL-6 in the wound area. The inflammatory proteins were decreased in tissues from the CCL4-knockout mice ((G); n = 3). WT, wild-type mice; CCL4KO, CCL4-knockout mice; WT-DM, wild-type diabetic mice; CCL4KO-DM, CCL4-knockout diabetic mice. * p < 0.05, ** p < 0.01 compared with WT, # p < 0.05 compared with WT-DM.

Article Snippet: Cells were transfected with CCL4 siRNA (Santa Cruz, sc-43932, Dallas, TX, USA) using Oligofectamine (Invitrogen, 12252011, Carlsbad, CA, USA) in opti-MEM.

Techniques: Inhibition, Knock-Out, Staining, Immunostaining, Western Blot

Journal: Biomedicines

Article Title: CCL4 Deletion Accelerates Wound Healing by Improving Endothelial Cell Functions in Diabetes Mellitus.

doi: 10.3390/biomedicines10081963

Figure Lengend Snippet: Figure 4. CCL4 inhibition by knockout enhanced neovascularization in diabetic mice. Representative Matrigel plug (A) and analysis of hemoglobin content ((B); n = 6). Representative images with H&E staining (C). Representative images with immunostaining of CD31 and Ki67. Both CD31- and Ki67-positive areas were enhanced in the CCL4-knockout mice (D,E). Serum concentrations of VEGF and SDF-1α were higher in the CCL4-knockout diabetic mice than in the wild-type diabetic mice ((F,G); n = 6). WT, wild-type mice; CCL4KO, CCL4-knockout mice; WT-DM, wild-type diabetic mice; CCL4KO-DM, CCL4-knockout diabetic mice. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were transfected with CCL4 siRNA (Santa Cruz, sc-43932, Dallas, TX, USA) using Oligofectamine (Invitrogen, 12252011, Carlsbad, CA, USA) in opti-MEM.

Techniques: Inhibition, Knock-Out, Staining, Immunostaining