Journal: Cell Reports

Article Title: Klebsiella pneumoniae hijacks the Toll-IL-1R protein SARM1 in a type I IFN-dependent manner to antagonize host immunity

doi: 10.1016/j.celrep.2022.111167

Figure Lengend Snippet:

Article Snippet: B16-Blue IFN-α/β reporter cells , InvivoGen , Cat# bb-ifnt1.

Techniques: Virus, Recombinant, Purification, Enzyme-linked Immunosorbent Assay, Software

Journal: Cardiovascular Therapeutics

Article Title: A Positive Feedback Loop Between CXCL16 and the Inflammatory Factors IL-17A and TGF- β Promotes Large Artery Atherosclerosis by Activating the STAT3/NF- κ B Pathway

doi: 10.1155/cdr/2973633

Figure Lengend Snippet: Primers used in this study.

Article Snippet: The membrane was blocked with milk (2.5 g milk powder in 50 mL double distilled water) at room temperature for 2 h. The cells were incubated with a primary antibody (diluted 1:1000, anti-CXCL16 antibody from Affinity) at 4°C for 8 h. Then, the membranes were incubated with secondary antibodies from Proteintech that were diluted at 1:2000 at room temperature for 2 h. Then, the protein bands on the membranes were visualized with white LED's bar Epi-IL fluorescence under the action of ECL hypersensitive chromogenic reagent (Yeasen, Shanghai, China).

Techniques:

Journal: Cardiovascular Therapeutics

Article Title: A Positive Feedback Loop Between CXCL16 and the Inflammatory Factors IL-17A and TGF- β Promotes Large Artery Atherosclerosis by Activating the STAT3/NF- κ B Pathway

doi: 10.1155/cdr/2973633

Figure Lengend Snippet: Assessment of CXCL16 expression in plaques by immunohistochemical staining. (a) Patient general characteristics. (b) Immunohistochemical analysis. Immunohistochemical staining of carotid plaque intima was performed using a CXCL16 antibody, and surface density (accumulated optical density value (IOD)/tissue area to be measured), histochemical score (H-score), and positive rate (number of positive cells/total number of cells) were analysed. The unpaired t -test was used to analyse the differences between the two groups, with all the data presented as the means ± SDs. n = 10; ⁣ ∗ p < 0.05; ⁣ ∗∗ p < 0.01. (c) The expression levels of CXCL16 in vulnerable plaques and stable plaques were compared by WB and RT–PCR. An unpaired t -test was used to analyse the differences between the two groups, with all data presented as the means ± SDs. n = 10, ⁣ ∗∗∗∗ p < 0.0001.

Article Snippet: The membrane was blocked with milk (2.5 g milk powder in 50 mL double distilled water) at room temperature for 2 h. The cells were incubated with a primary antibody (diluted 1:1000, anti-CXCL16 antibody from Affinity) at 4°C for 8 h. Then, the membranes were incubated with secondary antibodies from Proteintech that were diluted at 1:2000 at room temperature for 2 h. Then, the protein bands on the membranes were visualized with white LED's bar Epi-IL fluorescence under the action of ECL hypersensitive chromogenic reagent (Yeasen, Shanghai, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: Cardiovascular Therapeutics

Article Title: A Positive Feedback Loop Between CXCL16 and the Inflammatory Factors IL-17A and TGF- β Promotes Large Artery Atherosclerosis by Activating the STAT3/NF- κ B Pathway

doi: 10.1155/cdr/2973633

Figure Lengend Snippet: Macrophages cocultured with HUVECs after regulating the expression levels of IL-17A and TGF- β . (a) The expression levels of CXCL16, IL-17A, and TGF- β were analysed by ELISA. One-way ANOVA was used to analyse the differences between the groups, and the data are shown as means ± SDs. n = 6; ⁣ ∗ p < 0.05; ⁣ ∗∗ p < 0.01; ⁣ ∗∗∗ p < 0.001; ⁣ ∗∗∗∗ p < 0.0001. (b) The expression of CXCL16 mRNA was detected by RT–PCR. One-way ANOVA was used to analyse the differences between the groups, and the data are shown as means ± SDs. n = 3; ⁣ ∗ p < 0.05. (c, d) The expression levels of CXCL16, STAT3, p-STAT3, and NF- κ B were measured by WB. One-way ANOVA was used to analyse the differences between the groups, and the data are shown as means ± SDs. n = 3; ⁣ ∗ p < 0.05; ⁣ ∗∗ p < 0.01; ⁣ ∗∗∗ p < 0.001; ⁣ ∗∗∗∗ p < 0.0001.

Article Snippet: The membrane was blocked with milk (2.5 g milk powder in 50 mL double distilled water) at room temperature for 2 h. The cells were incubated with a primary antibody (diluted 1:1000, anti-CXCL16 antibody from Affinity) at 4°C for 8 h. Then, the membranes were incubated with secondary antibodies from Proteintech that were diluted at 1:2000 at room temperature for 2 h. Then, the protein bands on the membranes were visualized with white LED's bar Epi-IL fluorescence under the action of ECL hypersensitive chromogenic reagent (Yeasen, Shanghai, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Cardiovascular Therapeutics

Article Title: A Positive Feedback Loop Between CXCL16 and the Inflammatory Factors IL-17A and TGF- β Promotes Large Artery Atherosclerosis by Activating the STAT3/NF- κ B Pathway

doi: 10.1155/cdr/2973633

Figure Lengend Snippet: HUVECs with controlled expression of CXCL16 were cocultured with macrophages with controlled expression of IL-17A and TGF- β . (a–c) The expression levels of CXCL16, IL-17A, and TGF- β were determined by ELISA. One-way ANOVA was used to analyse the differences between the groups, and the data are shown as means ± SDs. Group A, n = 7.

Article Snippet: The membrane was blocked with milk (2.5 g milk powder in 50 mL double distilled water) at room temperature for 2 h. The cells were incubated with a primary antibody (diluted 1:1000, anti-CXCL16 antibody from Affinity) at 4°C for 8 h. Then, the membranes were incubated with secondary antibodies from Proteintech that were diluted at 1:2000 at room temperature for 2 h. Then, the protein bands on the membranes were visualized with white LED's bar Epi-IL fluorescence under the action of ECL hypersensitive chromogenic reagent (Yeasen, Shanghai, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cardiovascular Therapeutics

Article Title: A Positive Feedback Loop Between CXCL16 and the Inflammatory Factors IL-17A and TGF- β Promotes Large Artery Atherosclerosis by Activating the STAT3/NF- κ B Pathway

doi: 10.1155/cdr/2973633

Figure Lengend Snippet: HUVECs with controlled expression of CXCL16 were cocultured with macrophages with controlled expression of IL-17A and TGF- β . (a, b) The expression levels of CXCL16-, NF- κ B-, STAT3-, and p-STAT3-related factors were measured by WB, and the differences among the groups were analysed by one-way ANOVA. The data are shown as means ± SDs, n = 3.

Article Snippet: The membrane was blocked with milk (2.5 g milk powder in 50 mL double distilled water) at room temperature for 2 h. The cells were incubated with a primary antibody (diluted 1:1000, anti-CXCL16 antibody from Affinity) at 4°C for 8 h. Then, the membranes were incubated with secondary antibodies from Proteintech that were diluted at 1:2000 at room temperature for 2 h. Then, the protein bands on the membranes were visualized with white LED's bar Epi-IL fluorescence under the action of ECL hypersensitive chromogenic reagent (Yeasen, Shanghai, China).

Techniques: Expressing

Journal: Journal of Bacteriology

Article Title: Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species ▿ †

doi: 10.1128/JB.00101-10

Figure Lengend Snippet: Neisseria strains used in this study

Article Snippet: FA1090 (ST-1899) , , , , ATCC 700825.

Techniques: Glycoproteomics, Modification

Journal: Journal of Bacteriology

Article Title: Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species ▿ †

doi: 10.1128/JB.00101-10

Figure Lengend Snippet: Neisserial glycoprotein profiles and glycan diversity. (Top panels) Immunoblotting of whole-cell lysates from strains of N. gonorrhoeae (Ngo) (N400 pglC, N400, and FA1090), N. meningitidis (Nme) (MC58, H44/76, Z2491, 8013, FAM18, BZ 10, and BZ 198), and N. lactamica (Nla) (ST-3787 and ST-640) with glycan-specific monoclonal antibodies. (Bottom panels) Immunoblotting using the SM1 MAb specifically recognizing class I pilin types and a polyclonal antiserum raised against the PilE-derived peptide 44KSAVTEYYLNHGKWPENNTSA64, which reacts with all pilin types. Thus, strains FAM18, BZ 10, ST-3787, and ST-640 all express class II pilins. Also, strains 8013, FAM18, BZ 10, and BZ 198 carry the pglB2 allele that is associated with synthesis of GATDH glycans, while all of the other strains have the pglB allele that is associated with synthesis of DATDH glycans.

Article Snippet: FA1090 (ST-1899) , , , , ATCC 700825.

Techniques: Glycoproteomics, Western Blot, Bioprocessing, Derivative Assay

Journal: Journal of Bacteriology

Article Title: Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species ▿ †

doi: 10.1128/JB.00101-10

Figure Lengend Snippet: Glycoprotein profiles and glycan antigenicity in strains N400, FA1090, and MC58. (A) Whole-cell lysates of N. gonorrhoeae N400 and FA1090 wild-type (wt) strains and pglA (monosaccharide-expressing) and pglC (glycosylation-null) mutants probed with MAb npg1. The strains used were KS100 (N400 wt), KS104 (N400 pglC), KS141 (N400 pglA), KS302 (FA1090 pglA), KS301 (FA1090 pglC), and KS300 (FA1090 wt). (B) Immunoblotting of whole-cell lysates of N. gonorrhoeae FA1090 and N. meningitidis MC58 wild-type strains and pglA (monosaccharide-expressing) and pglC (glycosylation-null) mutants probed with MAbs npg1, npg2, and npg3. The strains used were KS301 (FA1090 pglC), KS302 (FA1090 pglA), KS300 (FA1090 wt), KS317 (MC58 pglC), KS318 (MC58 pglA), and KS316 (MC58 wt).

Article Snippet: FA1090 (ST-1899) , , , , ATCC 700825.

Techniques: Glycoproteomics, Expressing, Western Blot

Journal: Journal of Bacteriology

Article Title: Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species ▿ †

doi: 10.1128/JB.00101-10

Figure Lengend Snippet: Increased npg2 and npg3 immunoreactivity at high pH and in pglI mutants. Immunoblotting of whole-cell lysates of wild-type strains, pglI mutants, and high-pH-treated wild-type strains revealed increased reactivity with the npg2 and npg3 monoclonal antibodies due to the lack of an O-acetyl group in pglI mutants or with the high-pH treatment. The extraneous band that appeared during high-pH treatment is PilE (as determined by immunoblotting with SM1 [data not shown]). This band was not present with the 4/3/1 background, in which pilE was conditionally repressed. The strains used were KS100 (N400 wt), KS144 (N400 pglI), KS104 (N400 pglC), KS101 (4/3/1 wt), KS300 (FA1090 wt), KS303 (FA1090 pglI), and KS301 (FA1090 pglC). wt, wild type.

Article Snippet: FA1090 (ST-1899) , , , , ATCC 700825.

Techniques: Western Blot, Bioprocessing

Journal: Journal of Bacteriology

Article Title: Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species ▿ †

doi: 10.1128/JB.00101-10

Figure Lengend Snippet: Protein glycosylation genotypes and phenotypes

Article Snippet: FA1090 (ST-1899) , , , , ATCC 700825.

Techniques: Glycoproteomics