recombinant mouse ccl22  (MedChemExpress)

Journal: Cell Biology and Toxicology

Article Title: SENP3 mediated DeSUMOylation of macrophage derived CCL17 accelerates atherosclerosis via regulation of Treg

doi: 10.1007/s10565-025-10099-3

Figure Lengend Snippet: Macrophages stimulated by oxLDL affect the chemotaxis of Treg through CCL17 and CCL22 competition. Macrophages were stimulated with oxLDL, and Treg cells were co-cultured with macrophage supernatant. A Immunofluorescence detection of co-localization of CCL22 and macrophages (CD68) in plaque tissue of AS mice. B The content of CCL22 in the supernatant of macrophages was detected by ELISA. C Transwell detected the effect of macrophages on the chemotaxis of Treg. ( D - E ) Transwell detected the migration of Treg cells. * p < 0.05, ** p < 0.01, *** p < 0.001. N = 3

Article Snippet: Prior to the transwell migration assay, the induced Tregs were treated with recombinant mouse CCL17 (MCE, HY-P71891A) or recombinant mouse CCL22 (MCE, HY-P7248) or anti-CCL17 (R&D, Catalog #: MAB529) and anti-CCL22 antibodies (R&D, Catalog #: MAB529).

Techniques: Chemotaxis Assay, Cell Culture, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Migration

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A . Differentially expressed genes in unstimulated or LPS or IL-4 stimulated LysM ΔZeb1 compared to LysM Ctrl BMDMs as measured by a customized RT2 array and depicted in log 2 fold change of expression. nd marks non-detectable mRNA levels. All transcripts were normalized to Gapdh (n=3). B . Relative mRNA expression of Ccl2 and Ccl22 in LysM Ctrl and LysM ΔZeb1 BMDMs (n=3; means ±SD; 2-way ANOVA). C . Comparison of transcript and secretome alterations of Ccl2 and Ccl22 in LysM ΔZeb1 compared to LysM Ctrl BMDMs (n≥5; means ±SD).

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Expressing, Comparison

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A-B . Venn diagrams of differentially expressed genes (DEGs) of LysM Ctrl (blue) and LysM ΔZeb1 BMDMs (red) after stimulation with LPS (A) or IL-4 (B) compared to unstimulated in total (left) and divided in up-/downregulated DEGs (right). C . GO term enrichment analysis for DEGs (FDR<0.05) uniquely up- or downregulated by LysM ΔZeb1 BMDMs after LPS stimulation. D . Log 2 fold change of expression of selected trafficking genes after LPS stimulation. X marks no significant deregulation. E . Representative images and quantification of OPP incorporation of LysM Ctrl and LysM ΔZeb1 BMDMs with 0h, 4h and 16h LPS pre-stimulation (n=3; means ±SD; 2-way ANOVA). F-G . Representative arrays of intracellular cytokines of LysM Ctrl and LysM ΔZeb1 BMDMs with LPS stimulation or additional Brefeldin A and Monensin treatment (F) and quantification of intracellular CCL2 and CCL22 after LPS, Brefeldin A and Monensin treatment (G; n≥2).

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Expressing

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A . Confluence of KPC cells alone or co-cultured with LysM Ctrl or LysM ΔZeb1 BMDMs (n=3). B . Representative images at t=28h and quantification over time of KPC cell invasion into a scratch wound without or with co-culture of LysM Ctrl or LysM ΔZeb1 BMDMs (n=3). C . Confluence of KPC cells alone or with LysM Ctrl or LysM ΔZeb1 BMDM conditioned medium (CM) (n=2 KPC CM, n=3 LysM Ctrl and LysM ΔZeb1 CM). D . Transwell migration assay of CD8+ T cells alone or towards LysM Ctrl or LysM ΔZeb1 BMDMs in absence or presence of recombinant CCL2 and CCL22 (left panel, n>3) or absence or presence of anti-CCL2 and anti-CCL22 antibodies (right panel, n=3). Means ±SD; *:p<0.05; **:p<0.01; ns: not significant; 2-way ANOVA.

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Cell Culture, Co-Culture Assay, Transwell Migration Assay, Recombinant

Journal: bioRxiv

Article Title: CCR4 and CCR7 differentially regulate thymocyte subset localization with distinct outcomes for central tolerance

doi: 10.1101/2022.04.03.486911

Figure Lengend Snippet: A-D) Transwell assays were used to quantify chemotaxis of thymocyte subsets to the indicated concentrations of CCL22 (A), CCL21 (B), CCL17 (C) and CCL19 (D). Migration indexes were calculated as the frequencies of input cells of each subset that migrated towards the chemokine relative to the frequencies of input cells that migrated in the absence of chemokine (Blank). Data were compiled from 3 independent experiments with triplicate wells per assay. 2-way ANOVA with Dunnett’s multiple comparison correction were used for statistical analysis. (Mean ± SEM; *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001)

Article Snippet: The bottom chamber of the transwells contained recombinant mouse chemokines CCL17, CCL22, CCL19 or CCL21 (Peprotech) at 100nM, 10nM and 1nM, diluted in 500μL RPMI 1640 + 10% FBS.

Techniques: Chemotaxis Assay, Migration, Comparison

Journal: bioRxiv

Article Title: CCR4 and CCR7 differentially regulate thymocyte subset localization with distinct outcomes for central tolerance

doi: 10.1101/2022.04.03.486911

Figure Lengend Snippet: A) and B) Expression of CCR4 ligands (A) and CCR7 ligands (B) by distinct thymic antigen presenting cell subsets, assessed by qRT-PCR. Expression levels were normalized to subsets previously reported to expresses the respective ligands: CCL17 and CCL22 expression were normalized to cDC2 ( Hu et al ., 2015b ), and CCL19 and CCL21 expression to mTEClo ( ; ). Data were compiled from 3 independent experiments with triplicates. One-way ANOVA with Dunnett’s multiple comparison correction were used for statistical analysis. (Mean ± SEM; ns: p>0.05, **: p<0.01) C) Schematic of experiments to determine if Ccr4 -/- thymocytes proliferate in response to congenic APCs activated by TLR stimulation. D) The percentage of WT or Ccr4 -/- T cells that proliferated at day 3 and day 5 when cultured without splenocytes, with unstimulated splenocytes, or with LPS-stimulated splenocytes, as indicated. Data were compiled from 4 independent experiments with triplicate wells, and unpaired t-tests with Holm-Šídák’s multiple comparison correction was used for statistical analysis. (Mean ± SEM; *: p<0.05)

Article Snippet: The bottom chamber of the transwells contained recombinant mouse chemokines CCL17, CCL22, CCL19 or CCL21 (Peprotech) at 100nM, 10nM and 1nM, diluted in 500μL RPMI 1640 + 10% FBS.

Techniques: Expressing, Quantitative RT-PCR, Comparison, Cell Culture