murine tweak  (R&D Systems)

Journal: The FASEB Journal

Article Title: CD163 deficiency increases foam cell formation and plaque progression in atherosclerotic mice

doi: 10.1096/fj.202000177r

Figure Lengend Snippet: FIGURE 6 CD163 prevents pro-inflammatory actions induced by TWEAK. A, Representative western blot analysis of p-p65 and p65 in VSMCs treated with 0% FBS (control), rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL). Right panel show the quantification of band densitometry values of p-p65 protein levels expressed in arbitrary units after correction for p65 (loading control). *P < .01 vs Control; †P < .01 vs rTW; Student's t test. B, Relative CCL2, CCL5, ICAM-1, MMP-2, and MMP-9 mRNA expression levels normalized to 18S rRNA of VSMCs treated with rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL) during 24 hours. The same cDNA was repeatedly tested for multiple genes. Data represent the mean ± SEM of three independent experiments. *P < .05 vs Control; †P < .05 vs rTW. Student's t test. C, Effect of CD163 on CCL2 and CCL5 secretion induced by TWEAK in VSMCs. VSMCs were incubated with rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL) during 24 hours and supernatants were tested by ELISA for CCL2 or CCL5. Data represent the mean ± SEM of six independent experiments; *P < .001 vs control; †P < .001 vs rTW; Student's t test. D, Peritoneal macrophages were seeded in the upper surface of chemotaxis chambers and stimulated with the supernatants of VSMCs treated with 0% FBS (control), rTWEAK (100 ng/mL) or rTW plus rCD (0.1-0.5 μg/mL). Quantification of migrated cells in 10 fields per condition is shown in the right panel. Data represent the mean ± SEM of four independent experiments; *P < .001 vs Control; †P < .001 vs rTW; Student's t test. Scale bars, 50 μm

Article Snippet: For experimental analysis, cells were made quiescent by 24-hour incubation in medium without FBS before stimulation with recombinant murine TWEAK (0.1 μg/mL; 1237-TW; R&D Systems) and preincubated or not with different concentrations of rCD163 (7435-CD; R&D Systems).

Techniques: Western Blot, Control, Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Chemotaxis Assay

Journal: The FASEB Journal

Article Title: CD163 deficiency increases foam cell formation and plaque progression in atherosclerotic mice

doi: 10.1096/fj.202000177r

Figure Lengend Snippet: FIGURE 5 TWEAK enhances plaque instability in CD163 deficient mice. A, Representative photographs of α-SMA + DAPI and CD68 stained sections. Quantification of CD68 and α-SMA in aortic root from ApoE−/−CD163+/+ and ApoE−/−CD163−/− mice treated with rTW alone or rTW plus rCD in the right panel. Values shown are mean ± SEM of 8-9 animals per group. *P < .05 vs ApoE−/−CD163+/+; †P < .001 vs ApoE−/−CD163−/− + rTW; One-way ANOVA with Bonferroni's posttest. Scale bar, 100 μm. B, Percentage of each grade from Stary method between different groups. N = 20-25 plaques per group. C, Relative CCL2 and CCL5 mRNA expression levels normalized to 18S rRNA of aortas from ApoE−/−CD163+/+ or ApoE−/−CD163−/− mice treated with rTW alone or rTW plus rCD. Data represent the mean ± SEM of six animals per group; *P < .001 vs ApoE−/−CD163+/+ + rTW. †P < .001 vs ApoE−/−CD163−/− + rTW; One-way ANOVA with Bonferroni's posttest. D, Representative images of BCA cross sections stained with MOVAT from ApoE−/−CD163+/+ or ApoE−/−CD163−/− mice treated with rTW alone or rTW plus rCD. Dashed lines show the boundary of the developing necrotic area (NC). Quantification of the necrotic core area is shown in the below panel. Data represent the mean ± SEM of eight animals per group; *P < .05 vs ApoE−/−CD163+/+; †P < .001 vs ApoE−/−CD163−/− + rTW; One-way ANOVA with Bonferroni's posttest. Scale bar, 100 μm. E, LDH activity in serum from ApoE−/−CD163+/+ and ApoE−/−CD163−/− mice treated with rTW alone or rTW and rCD. Data represent the mean ± SEM of six animals per group; *P < .001, vs ApoE−/−CD163+/+ + rTW. †P < .001 vs ApoE−/−CD163−/− + rTW. One-way ANOVA with Bonferroni's posttest. F, Frequency of morphological markers of plaque instability in ApoE−/−CD163+/+ or ApoE−/−CD163−/− mice treated with rTW alone or rTW plus rCD. Data represent the mean ± SEM of eight animals per group; *P < .05 vs ApoE−/−CD163+/+ + rTW. †P < .001 vs ApoE−/−CD163−/− + rTW; One-way ANOVA with Bonferroni's posttest. G, Representative images of BCA cross sections stained with α-SMA or CD68 from ApoE−/−CD163+/+ or ApoE−/−CD163−/− mice treated with rTW alone or rTW plus rCD. Quantification CD68 positive cells is shown in the below panel. Data represent the mean ± SEM of eight animals per group; *P < .05 vs ApoE−/−CD163+/+ + rTW; †P < .001 vs ApoE−/−CD163−/− + rTW; One-way ANOVA with Bonferroni's posttest. Scale bar, 100 μm

Article Snippet: In addition, TWEAKaccelerated atherosclerosis was induced in 12-week-old ApoE−/−CD163-/- (N = 10), and ApoE−/−CD163-/- (N = 10) feeding a hyperlipidemic diet by murine TWEAK injection (5 μg/kg/three times per week; 1237-TW; R&D Systems) during the last 4 weeks.

Techniques: Staining, Expressing, Activity Assay

Journal: The FASEB Journal

Article Title: CD163 deficiency increases foam cell formation and plaque progression in atherosclerotic mice

doi: 10.1096/fj.202000177r

Figure Lengend Snippet: FIGURE 6 CD163 prevents pro-inflammatory actions induced by TWEAK. A, Representative western blot analysis of p-p65 and p65 in VSMCs treated with 0% FBS (control), rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL). Right panel show the quantification of band densitometry values of p-p65 protein levels expressed in arbitrary units after correction for p65 (loading control). *P < .01 vs Control; †P < .01 vs rTW; Student's t test. B, Relative CCL2, CCL5, ICAM-1, MMP-2, and MMP-9 mRNA expression levels normalized to 18S rRNA of VSMCs treated with rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL) during 24 hours. The same cDNA was repeatedly tested for multiple genes. Data represent the mean ± SEM of three independent experiments. *P < .05 vs Control; †P < .05 vs rTW. Student's t test. C, Effect of CD163 on CCL2 and CCL5 secretion induced by TWEAK in VSMCs. VSMCs were incubated with rTW (100 ng/mL) or rTW plus rCD (0.01-0.5 μg/mL) during 24 hours and supernatants were tested by ELISA for CCL2 or CCL5. Data represent the mean ± SEM of six independent experiments; *P < .001 vs control; †P < .001 vs rTW; Student's t test. D, Peritoneal macrophages were seeded in the upper surface of chemotaxis chambers and stimulated with the supernatants of VSMCs treated with 0% FBS (control), rTWEAK (100 ng/mL) or rTW plus rCD (0.1-0.5 μg/mL). Quantification of migrated cells in 10 fields per condition is shown in the right panel. Data represent the mean ± SEM of four independent experiments; *P < .001 vs Control; †P < .001 vs rTW; Student's t test. Scale bars, 50 μm

Article Snippet: In addition, TWEAKaccelerated atherosclerosis was induced in 12-week-old ApoE−/−CD163-/- (N = 10), and ApoE−/−CD163-/- (N = 10) feeding a hyperlipidemic diet by murine TWEAK injection (5 μg/kg/three times per week; 1237-TW; R&D Systems) during the last 4 weeks.

Techniques: Western Blot, Control, Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Chemotaxis Assay

Journal: Cell Death & Disease

Article Title: Fn14 deficiency ameliorates psoriasis-like skin disease in a murine model

doi: 10.1038/s41419-018-0820-6

Figure Lengend Snippet: Psoriasis-like disease was induced by imiquimod cream in the back of mice. Recombinant TWEAK (20 µg/ml, 100 μl each time, twice daily) was also topically applied to skin lesion accordingly. Skin tissue was harvested on day 7. a , b The HE-stained sections were evaluated by HPSS method. There were no significant differences between the two control groups of the WT mice or between the three groups of the KO mice. c By immunohistochemistry, the sections were detected for the Iba-1- or CD3-positive cells. d , e The positive cells were counted according to the areas of epidermis and dermis. Representative images are shown; n = 6 in each group. Bar = 20 μm

Article Snippet: In some experiments, recombinant murine TWEAK (R&D Systems, Minneapolis, MN, USA) or BSA (Sigma-Aldrich, St Louis, MO) was prepared in normal saline (20 µg/ml), and then twice daily (100 μl each time) administrated to the surface of the skin lesion.

Techniques: Cream, Recombinant, Staining, Immunohistochemistry