rabbit anti mouse traf6  (Proteintech)

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Corilagin relieves atherosclerosis via the toll-like receptor 4 signaling pathway in vascular smooth muscle cells

doi: 10.1177/03946320241254083

Figure Lengend Snippet: Primers of qRT-PCR.

Article Snippet: The primary antibodies used were rabbit anti-mouse TLR4 1:1000 (Cat No. 30400-1-AP, Proteintech, Beijing, China), rabbit anti-mouse MyD88 1:800 (Cat No. A0980, ABclonal), rabbit anti-mouse TIRAP 1:800 (Cat No. A12606, ABclonal), rabbit anti-mouse TRAF6 1:1000 (Cat No. 12809-1-AP, Proteintech), rabbit anti-mouse NEMO 1:1000 (Cat No. 18474-1-AP, Proteintech), rabbit anti-mouse p38MAPK 1:1000 (Cat No. 14064-1-AP, Proteintech), and rabbit anti-mouse IRF5 1:1000 (Cat No. 10547-1-AP, Proteintech).

Techniques:

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Corilagin relieves atherosclerosis via the toll-like receptor 4 signaling pathway in vascular smooth muscle cells

doi: 10.1177/03946320241254083

Figure Lengend Snippet: Effect of corilagin on the TLR4 signaling pathway in MOVAS cells stimulated by ox-LDL. (a) mRNA expression of TLR4, TIRAP, MyD88, TRAF6, p38, NEMO and IRF5 was measured by qRT-PCR. * p < .05 compared with the control group, # p < .05 compared with the ox-LDL group, ** p < .05 compared with the aspirin group as determined by one-way ANOVA and subsequent Student–Newman–Keuls q -test ( n = 5). (B. C) Protein abundance was measured by western blotting. * p < .05 compared with the control group, # p < .05 compared with the ox-LDL group, ** p < .05 compared with the aspirin group, as determined by one-way ANOVA and subsequent Student–Newman–Keuls q -test ( n = 5). (D, E) Abundance of IL-6 and MCP-1 in cell culture supernatant was measured by ELISAs. * p < .05 compared with the control group, # p < .05 compared with the ox-LDL group, ** p < .05 compared with the aspirin group as determined by one-way ANOVA and subsequent Student–Newman–Keuls q -test ( n = 5). (F) Effect of corilagin on the MOVAS cells proliferation. * p < .05 compared with the control group, # p < .05 compared with the ox-LDL group determined by one-way ANOVA and subsequent Student–Newman–Keuls q -test ( n = 5). (G, H) apoptosis ratio of MOVAS were detected by Annexin V staining. No significant difference was found among four groups determined by one-way ANOVA and subsequent Student–Newman–Keuls q -test ( n = 5).

Article Snippet: The primary antibodies used were rabbit anti-mouse TLR4 1:1000 (Cat No. 30400-1-AP, Proteintech, Beijing, China), rabbit anti-mouse MyD88 1:800 (Cat No. A0980, ABclonal), rabbit anti-mouse TIRAP 1:800 (Cat No. A12606, ABclonal), rabbit anti-mouse TRAF6 1:1000 (Cat No. 12809-1-AP, Proteintech), rabbit anti-mouse NEMO 1:1000 (Cat No. 18474-1-AP, Proteintech), rabbit anti-mouse p38MAPK 1:1000 (Cat No. 14064-1-AP, Proteintech), and rabbit anti-mouse IRF5 1:1000 (Cat No. 10547-1-AP, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Control, Quantitative Proteomics, Western Blot, Cell Culture, Staining

Journal: Neuroimmunomodulation

Article Title: MiR-124 Reduced Neuroinflammation after Traumatic Brain Injury by Inhibiting TRAF6.

doi: 10.1159/000528502

Figure Lengend Snippet: Fig. 5. Upregulation of miR-124 inhibited the activation of TLR4 pathway after TBI. a–e TBI increased the expression of TLR4 and its downstream molecules MyD88/TRAF6/IRAK1/NF-κB p65. MiR-124 mimics reduced the expression of these proteins, but miR-124 inhibitors produced the contrary effects. n = 12 in sham group, n = 12 in other three groups. p p < 0.05 versus sham group, #p < 0.05 versus TBI group.

Article Snippet: Equivalent amount of protein (40 μg) was loaded and separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membrane at 4°C for 50 min. Membranes were blocked with 5% nonfat milk solution in Tris-buffered saline with 0.1% Triton X-100 (TBST) for 1 h and then incubated overnight at 4°C with appropriate primary antibodies as follows: rabbit anti-mouse TLR4 antibody (Thermo Fisher, USA), rabbit anti-mouse myeloid differentiation factor 88 (MyD88) antibody (Santa Cruz, USA), rabbit anti-mouse TNFRassociated factor (TRAF6) antibody (Novus Biologicals, USA), rabbit anti-mouse NF-κB p65 antibody (GeneTex, USA), and rabbit anti-β-actin antibody (Proteintech, USA).

Techniques: Activation Assay, Expressing, Produced

Journal: Neuroimmunomodulation

Article Title: MiR-124 Reduced Neuroinflammation after Traumatic Brain Injury by Inhibiting TRAF6.

doi: 10.1159/000528502

Figure Lengend Snippet: Fig. 6. MiR-124 targeted TRAF6. a, b LPS elevated the expression of TRAF6 in microglial cells, upregulation of miR-124 declined the expression of TRAF6, while downregulation of miR-124 elevated the expression of TRAF6. c MiR-124 and TRAF6 had the latent binding sites. d The dual-luciferase reporter activity examina- tions of the targeting of miR-124 with TRAF6. e RNA-immunoprecipitation test of the enrichment of miR-124 with TRAF6 in Ago2 magnetic beads. n = 6 in each group, p p < 0.05 versus control group, #p < 0.05 versus LPS group.

Article Snippet: Equivalent amount of protein (40 μg) was loaded and separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membrane at 4°C for 50 min. Membranes were blocked with 5% nonfat milk solution in Tris-buffered saline with 0.1% Triton X-100 (TBST) for 1 h and then incubated overnight at 4°C with appropriate primary antibodies as follows: rabbit anti-mouse TLR4 antibody (Thermo Fisher, USA), rabbit anti-mouse myeloid differentiation factor 88 (MyD88) antibody (Santa Cruz, USA), rabbit anti-mouse TNFRassociated factor (TRAF6) antibody (Novus Biologicals, USA), rabbit anti-mouse NF-κB p65 antibody (GeneTex, USA), and rabbit anti-β-actin antibody (Proteintech, USA).

Techniques: Expressing, Binding Assay, Luciferase, Activity Assay, RNA Immunoprecipitation, Magnetic Beads, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: TRAF6 shows high expression in fatty liver caused by HFD (A) The statistical plot for body weight, liver weight, and liver index in chow-fed mice and HFD-fed mice. (B) The scatterplot for FPG, FINS, and HOMA-IR in chow-fed mice and HFD-fed mice. (C) H&E staining for liver tissues in chow-fed mice and HFD-fed mice (400×). (D) Oil red O staining for fatty liver tissues in chow-fed mice and HFD-fed mice (200×). (E) The scatterplot for the contents of TG and TC in liver tissues of chow-fed and HFD-fed mice. (F) Histogram representing TNF-α levels in serum of chow-fed mice and HFD-fed mice. (G) Western blot analysis of CPT1 protein expression in liver tissues of chow-fed and HFD-fed mice and its statistical diagram. (H) Filipin staining for the fatty liver cell model (200×) and the quantitative histogram. (I) Nile red staining for the fatty liver cell model (200×) and the statistical histogram. (J) Histogram of FFA content in liver cells after OA/PA induction. (K) The histogram for the content of TG and TC in the fatty liver cell model. (L) The expression of TRAF6 in fatty liver tissues of chow-fed and HFD-fed mice as detected by immunohistochemistry (400×). (M) qRT-PCR determination of TRAF6 expression in fatty liver tissues of chow-fed mice and HFD-fed mice and the statistical histogram. (N) The histogram for TRAF6 expression in the fatty liver cell model as detected by qRT-PCR. (O) The protein expression of TRAF6 in the fatty liver cell model as detected by western blot analysis. These data were measurement data, expressed as mean ± standard deviation. Data between two groups were compared utilizing independent sample t test, while data among multiple groups at variant time points were analyzed through repeated-measures ANOVA, followed by Bonferroni post hoc test. Compared with chow or control, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. In chow-fed mice and HFD-fed mice, each n = 10. In hepatocytes used as control and induced with OA/PA, each n = 3.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Expressing, Staining, Western Blot, Immunohistochemistry, Quantitative RT-PCR, Standard Deviation, Variant Assay, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: Silencing of TRAF6 inhibits the cholesterol accumulation in hepatocytes (A) The silencing efficiency of TRAF6 as detected by qRT-PCR. (B) The silencing efficiency of TRAF6 as detected by western blot analysis. (C) Filipin staining for hepatocytes treated with sh-NC or sh-TRAF6 and the quantitative histogram. (D) Nile red staining results and the statistical histogram. (E) Histogram of free FFA content in hepatocytes treated with sh-NC or sh-TRAF6. (F) The histogram for the contents of TG and TC in hepatocytes treated with sh-NC or sh-TRAF6. These data were measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared using one-way ANOVA followed by a Tukey’s test. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared with sh-NC. The cell experiment was repeated three times. In chow-fed mice and HFD-fed mice, each n = 10. In hepatocytes used as control and induced with OA/PA, each n = 3.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Quantitative RT-PCR, Western Blot, Staining, Standard Deviation, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: TRAF6 mediates ubiquitination of EZH2 (A) EZH2 expression in liver tissues of HFD-fed mice and chow-fed mice as detected by immunohistochemistry. (B) EZH2 expression after OA/PA induction as detected by qRT-PCR. (C) EZH2 expression after OA/PA induction as detected by western blot analysis. (D) The histogram for TRAF6 and EZH2 expression as detected by western blot analysis. (E) The interaction of TRAF6 with EZH2 as detected by coIP. (F) The level of EZH2 ubiquitination as detected by in vivo ubiquitination assay. These data were measurement data, expressed as mean ± standard deviation. Data between two groups were compared utilizing independent sample t test, while data among multiple groups were compared with the use of one-way ANOVA, followed by a Tukey’s test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus control or sh-NC. The cell experiment was repeated three times. In tissue experiment, n = 10.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Ubiquitin Proteomics, Expressing, Immunohistochemistry, Quantitative RT-PCR, Western Blot, In Vivo, Standard Deviation, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: TRAF6 promotes the expression of miR-429 and cholesterol accumulation in hepatocytes through ubiquitination of EZH2 (A) The silencing efficiency of EZH2 as detected by qRT-PCR. (B) The silencing efficiency of EZH2 as detected by western blot analysis. (C) The expression of TRAF6 and EZH2 after different treatment as detected by western blot analysis. (D) The expression of miR-429 after different treatment as detected by qRT-PCR. (E) The expression of TRAF6 and EZH2 after different treatment as detected by western blot analysis. (F) The expression of miR-429 after different treatment as detected by qRT-PCR. (G) Filipin staining for hepatocytes after different treatment. (H) Nile red staining for hepatocytes. (I) Statistical histogram of FFA content in the cells after different treatment. (J) The histogram for the content of TG and TC in hepatocytes after different treatment. These data were measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared with the use of one-way ANOVA, followed by Tukey’s test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus sh-NC or sh-NC + mimic-NC. #p < 0.05, ##p < 0.01, ###p < 0.001 versus sh-TRAF6 + sh-NC or sh-TRAF6 + mimic-NC. The cell experiment was repeated three times.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Expressing, Ubiquitin Proteomics, Quantitative RT-PCR, Western Blot, Staining, Standard Deviation

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: TRAF6 promotes cholesterol accumulation in hepatocytes through the EZH2/miR-429/PPARα axis (A) The expression of TRAF6, EZH2, and PPARα after different treatment as detected by western blot analysis. (B) The expression of miR-429 after different treatment as detected by qRT-PCR. (C) Filipin staining for hepatocytes after different treatment. (D) Nile red staining after different treatment. (E) Statistical histogram of FFA in the cells after different treatment. (E) The histogram for FFA levels in the cells. (F) The histogram for the content of TG and TC in hepatocytes after different treatment. These data were measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared with the use of one-way ANOVA, followed by a Tukey’s test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus sh-NC. #p < 0.05, ##p < 0.01, ###p < 0.001 versus sh-TRAF6 + sh-NC. The cell experiment was repeated three times.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Staining, Standard Deviation

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: TRAF6 promotes the occurrence of fatty liver through the EZH2/miR-429/PPARα axis (A) The expression of TRAF6, EZH2, and PPARα after different treatment as detected by western blot analysis. (B) The expression of miR-429 after different treatment as detected by qRT-PCR. (C) The statistical plot for body weight, liver weight, and liver index in mice with different treatment. (D) H&E staining for liver tissues in mice with different treatment (200×). (E) Oil red O staining for fatty liver tissues in mice with different treatment (200×). (F) The scatterplot for the content of TG and TC in liver tissues of chow-fed mice and HFD-fed mice. These data were measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared with the use of one-way ANOVA, followed by a Tukey’s test. ∗p < 0.05 versus chow-fed mice. ###p < 0.001 versus sh-NC. $$$p < 0.001 versus sh-TRAF6 + sh-NC. n = 10.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Staining, Standard Deviation

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: Comparison of lipid metabolism related indexes in mice with different treatments

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Comparison

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: HFD increases the expression of TRAF6 TRAF6 reduces the expression of EZH2 by enhancing the ubiquitination of EZH2, which inhibits H3K27me3 to promote miR-429 expression. miR-429 inhibits the expression of PPARα in the cytoplasm, leading to the development of fatty liver.

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Expressing, Ubiquitin Proteomics

Journal: Molecular Therapy. Nucleic Acids

Article Title: HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis

doi: 10.1016/j.omtn.2021.01.026

Figure Lengend Snippet: Primer sequences for qRT-PCR

Article Snippet: China) at room temperature for 20 min, the sections were incubated overnight at 4°C with diluted primary rabbit anti-mouse antibodies to TRAF6 (ab33915, 1:1,000), EZH2 (5246, 1:300, Cell Signaling Technology, Beverly, MA, USA), and PPARα (ab215270, 1:1,000).

Techniques: Sequencing