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: PLoS ONE

Article Title: LncRNA HOTAIR promotes proliferation, invasion and migration in NSCLC cells via the CCL22 signaling pathway

doi: 10.1371/journal.pone.0263997

Figure Lengend Snippet: A, B: the expression of HOTAIR and CCL22 mRNA were detected by RT-qPCR. C: CCL22 protein analyzed by ELISA assay. ****, P<0.001.

Article Snippet: The concentration of CCL22 in the supernatant was tested according to the instructions of ELISA kit operation (CUSABIO, CSB-E04660h).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: LncRNA HOTAIR promotes proliferation, invasion and migration in NSCLC cells via the CCL22 signaling pathway

doi: 10.1371/journal.pone.0263997

Figure Lengend Snippet: A: The HOTAIR and CCL22 mRNA expression examined by RT-qPCR B: Cell apoptosis detected by Flow cytometry. C: The cell proliferation tested by MTS assay. D: The CCL22 protein expression tested by ELISA. E: The cell migration and invasion detected by Transwell assay. **, P<0.05; ***, P<0.01; ****, P<0.001;ns, no significance.

Article Snippet: The concentration of CCL22 in the supernatant was tested according to the instructions of ELISA kit operation (CUSABIO, CSB-E04660h).

Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry, MTS Assay, Enzyme-linked Immunosorbent Assay, Migration, Transwell Assay

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Analysis of cis -element in the Ccl22 promoter. JAWSII cells ( a ) and RAW264.7 cells ( b ) were transfected with reporter plasmids for renilla luciferase as an internal control and with reporter plasmids for firefly luciferase containing various lengths of the mouse Ccl22 promoter. ( c ) Sequences of the −27/−1 region of the mouse Ccl22 promoter. Two Ets motifs, designated Site1 and Site 2, are indicated in bold. ( d ) JAWSII cells were transfected with reporter plasmids of WT or mutant promoters lacking the Ets motif(s) at the indicated sites. All results are shown as means + S.D.s ( n = 3). Similar results were obtained in three independent experiments. * p < 0.05.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Transfection, Luciferase, Control, Mutagenesis

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Effects of PU.1 knockdown on Ccl22 expression in mouse DCs and macrophages. JAWSII ( a ), RAW264.7 ( b ), BMDCs ( c , e ), and BMDMs ( d and f ) were transfected with either negative control siRNA (siNega) or Spi1 siRNA (siSpi1). At 48 h after transfection, relative mRNA levels were determined by quantitative RT-PCR after normalizing to mouse Gapdh mRNA levels. Data are expressed as the ratio of the expression level of the respective control siRNA-transfected cells. Western blotting analyses using anti-PU.1 Ab and anti-β-actin Ab were performed to evaluate the effect of Spi1 siRNA on PU.1 protein levels in BMDCs ( c right) and BMDMs ( d right). ( e , f ) The concentration of the CCL22 protein produced from either siNega or siSpi1 transfected BMDCs ( e ) or BMDMs ( f ) was determined as described in the Materials and Methods . Results are shown as means + S.D.s ( n = 3). Similar results were obtained in three independent experiments. * p < 0.05.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Knockdown, Expressing, Transfection, Negative Control, Quantitative RT-PCR, Control, Western Blot, Concentration Assay, Produced

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Analysis of the PU.1 binding region in the mouse Ccl22 promoter. ChIP assay was performed using either goat IgG (gIgG) or anti-PU.1 Ab (PU.1) in BMDCs ( a ), BMDMs ( b ), splenic DCs ( c ), and peritoneal macrophages ( d ). The amounts of immunoprecipitated chromatin were determined by quantitative PCR amplifying the indicated region of the Ccl22 promoter. Data are expressed as the percentage of the input for each ChIP assay. Results are shown as means + S.D.s ( n > 3 ). Similar results were obtained in more than three independent experiments. * p < 0.05.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Binding Assay, Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Involvement of IRFs in the expression of Ccl22 in BMDCs. ( a , b ) BMDCs were transfected with negative control siRNA (siNega), Irf4 siRNA (siIrf4) ( a ), or Irf8 siRNA (siIrf8) ( b ). At 48 h after transfection, relative mRNA levels were determined by quantitative RT-PCR after normalizing to mouse Gapdh mRNA levels. Data are expressed as the ratio of the expression level of the respective control siRNA-transfected cells. Western blotting analyses were performed using transfectants, which were harvested at 48 h after transfection (right in a , b ). ( c ) ChIP assay was performed by using either goat IgG (gIgG) or anti-IRF4 Ab (IRF4) in BMDCs. The amounts of immunoprecipitated chromatin were determined by quantitative PCR targeting the −17/+52 region of the Ccl22 promoter. Data are expressed as the percentage of the input for each ChIP assay. ( d ) mRNA level (left) and protein level (right) of PU.1 in control (siNega) or Irf4 knockdown cells (siIrf4). ( e ) PU.1 binding degree to −17/+52 in control (siNega) or Irf4 knockdown cells (siIrf4) determined by ChIP assay. Results are shown as means + S.D.s ( n = 3 ). Similar results were obtained in three independent experiments. * p < 0.05.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Control, Western Blot, Immunoprecipitation, Real-time Polymerase Chain Reaction, Knockdown, Binding Assay

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Involvement of PU.1 in the expression of Ccl22 in mature BMDCs. ( a ) BMDCs were stimulated with 1 μg/ml LPS for the indicated period. Relative Ccl22 and Ccl17 mRNA levels were determined by quantitative RT-PCR after normalizing to mouse Gapdh mRNA levels. Data are expressed as the ratio of the expression level of the non-stimulated cells. ( b ) ChIP assay was performed using either goat IgG (gIgG) or anti-PU.1 Ab (α-PU.1) in BMDCs stimulated with 1 μg/ml LPS for 24 hours. The amounts of immunoprecipitated chromatin were determined by quantitative PCR targeting the −17/+52 region of the Ccl22 promoter. Data are expressed as the ratio of α-PU.1 to gIgG (α-PU.1/gIgG). ( c ) BMDCs were transfected with either negative control siRNA (siNega) or Spi1 siRNA (siSpi1). At 24 hours after transfection, cells were stimulated with 1 μg/ml LPS and further incubated for 24 hours. Relative mRNA levels were determined by quantitative RT-PCR after normalizing to mouse Gapdh mRNA levels. Data are expressed as the ratio of the expression level of the control siRNA-transfected immature BMDCs. Results are shown as means + S.D.s ( n > 3 ). Similar results were obtained in more than three independent experiments.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Expressing, Quantitative RT-PCR, Immunoprecipitation, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Incubation, Control

Journal: Scientific Reports

Article Title: A transcription factor PU.1 is critical for Ccl22 gene expression in dendritic cells and macrophages

doi: 10.1038/s41598-018-37894-9

Figure Lengend Snippet: Involvement of PU.1 in the expression of the human CCL22 gene. ( a ) mRNA levels of CCL22 and SPI1 in SPI1 siRNA (siSPI1) or control siRNA (siNega) introduced THP-1 cells. ( b ) Alignment of nucleotide sequences of human and mouse CCL22 genes. ( c ) ChIP assay was performed using either control IgG (gIgG) or anti-PU.1 Ab (α-PU.1) in THP-1 cells. The amounts of immunoprecipitated chromatin were determined by quantitative PCR targeting the −153/−76 (including cis -elements) or −2106/−2046 ( cis -control) of the CCL22 promoter. Results are shown as means + S.D.s of three independent experiments.

Article Snippet: The concentration of mouse CCL22 protein was determined using an ELISA kit (MCC220, R&D systems, Minneapolis, MN).

Techniques: Expressing, Control, Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Cell reports

Article Title: The IRAK1/IRF5 axis initiates IL-12 response by dendritic cells and control of Toxoplasma gondii infection

doi: 10.1016/j.celrep.2024.113795

Figure Lengend Snippet: (A) Volcano plot depicting T. gondii -infected versus uninfected CD11c + DCs (WT). The horizontal dashed line represents a —log false discovery rate of 2, and the vertical dashed lines represent a log 2 fold change of −1 and +1. (B) Heatmap showing the expression and hierarchical clustering of selected genes in CD11c + DCs from the indicated strains, uninfected or infected with T. gondii Me49 tachyzoites (MOI 3, 4 h). (C–F) Quantification of IL-12 p40 (C), CCL5 (D), TNF (E), and CCL22 (F) in culture supernatants of CD11c + DCs of the indicated strains infected with T. gondii Me49 (MOI 3) for 4 and 24 h. In (A) and (B), Data were pooled from four independent experiments. In (C)–(F), data were pooled from four independent experiments and are presented as mean ± SEM; each independent experiment consisted of one technical replicate. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (ANOVA with Tukey’s multiple-comparisons test).

Article Snippet: Cytokine and chemokine quantification was performed from culture supernatants, peritoneal lavage or serum, according to the manufacturer instructions: IL-12 p40 Mouse uncoated ELISA kit (Invitrogen, 88–7120-88), Mouse CCL5/Rantes DuoSet ELISA (R&D Systems, DY478), Mouse TNF-alpha DuoSet ELISA (R&D Systems, DY410), Mouse Interferon-gamma DuoSet ELISA (R&D Systems, DY485), and Mouse CCL22 DuoSet ELISA (R&D Systems, DY439).

Techniques: Infection, Expressing

Journal: Cell reports

Article Title: The IRAK1/IRF5 axis initiates IL-12 response by dendritic cells and control of Toxoplasma gondii infection

doi: 10.1016/j.celrep.2024.113795

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cytokine and chemokine quantification was performed from culture supernatants, peritoneal lavage or serum, according to the manufacturer instructions: IL-12 p40 Mouse uncoated ELISA kit (Invitrogen, 88–7120-88), Mouse CCL5/Rantes DuoSet ELISA (R&D Systems, DY478), Mouse TNF-alpha DuoSet ELISA (R&D Systems, DY410), Mouse Interferon-gamma DuoSet ELISA (R&D Systems, DY485), and Mouse CCL22 DuoSet ELISA (R&D Systems, DY439).

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, Marker, Western Blot, Extraction, Enzyme-linked Immunosorbent Assay, Software

Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: ( A ) GO analysis of macrophage response to LNR. ( B ) Heatmaps of macrophage-secreted chemokines in the iWAT from mice receiving sham or LNR at 6°C for 7 days ( n = 4 per group). Red and blue represent the fold increase and decrease in a gene, respectively (see color scale). ( C ) mRNA expression of macrophage-secreted chemokines in iWAT from mice described in (B) ( n = 8 per group). ( D ) Ccl22 mRNA expression in iWAT from mice described in (B) at 23° or 6°C for 7 days ( n = 6 per group). ( E ) Ccl22 mRNA expression in iWAT from mice described in (B) exposed to 6°C for different hours ( n = 6). ( F ) Serum CCL22 levels from mice described in (B) exposed to 6°C for different days ( n = 6). ( G ) Serum CCL22 levels from mice described in (B) ( n = 6 per group). ( H ) Ccl22 mRNA expression in iWAT SVF cells or mature adipocytes (MAs). ( I ) Ccl22 mRNA expression in M0, M1, and M2 macrophages derived from 10-week-old C57BL/6 bone marrows ( n = 8 per group). ( J and K ) mRNA expression (J) and IF (K) of UCP1 in beige adipocytes ( n = 6 per group). Scale bars, 25 μm. ( L to N ) Immunofluorescence of UCP1 (L), mRNA expression of thermogenic genes (M), and immunoblots of UCP1 (N) in iWAT ( n = 3 to 6 per group). Scale bars, 50 μm. ( O to R ) Immunoblots and quantification of UCP1 [(O) and (P)], mRNA expression (Q) of thermogenic genes, and IF of UCP1 and H&E staining (R) in iWAT ( n = 3 to 6 per group). Scale bars, 50 μm. Data information: Results are presented as means ± SEM. [(C), (D), (G) to (J), (M), (P), and (Q)] * P ≤ 0.05, ** P < 0.01, and *** P < 0.001 by nonpaired Student’s t test. [(E) and (F)] * P ≤ 0.05 and ** P < 0.01 nonpaired Student’s t test compared with before cold stimulation.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Expressing, Derivative Assay, Immunofluorescence, Western Blot, Staining

Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: ( A ) Pearson’s correlation coefficient analysis of serum CCL22 concentration and fat mass ( n = 28 per group). ( B to F ) Body weight (B), energy consumption rate [(C) and (D)], and RER [(E) and (F)] ( n = 8 per group). Ten-week-old C57BL/6 male mice received saline or rCCL22 (20 μg/kg per day) at 6°C for 14 days with HFD. ( G to I ) Immunofluorescence (G), immunoblots (H) of UCP1, and mRNA expression (I) of thermogenic genes in iWAT ( n = 3 to 6 per group). Scale bar, 50 μm. ( J to L ) Immunofluorescence (J), immunoblots, and quantification [(K) and (L)] of UCP1 in iWAT ( n = 3 per group). SVF cells were extracted from HFD-fed iWAT, cultured with vehicle or rCCL22 (10 ng/ml) for 4 days, and then induced to beige adipocytes for 5 days. Scale bars, 25 μm. ( M ) Schematic diagram: The 6-month trial of obese adults participated in ADF: a 3-month weight loss period followed by a 3-month weight maintenance period. ( N and O ) Pearson’s correlation coefficient analysis of human serum CCL22 levels, body weight (N), and fat mass (O) ( n = 40). ( P ) Human serum CCL22 levels ( n = 20). ( Q ) Schematic diagram: Human SVFs from iWAT were cultured at 31°C for 5 days, treated with vehicle or CCL22 (10 ng/ml) for 4 days, and then induced to beige adipocytes. ( R to T ) Immunoblots and quantification [(R) and (S)] of UCP1 and mRNA expression (T) of thermogenic genes in human beige adipocytes ( n = 3 to 6 per group). Data information: Results are presented as means ± SEM. [(A), (N), and (O)] Two-tailed Pearson’s correlation coefficient analysis. [(F), (I), (L), (S), and (T)] * P ≤ 0.05 and ** P < 0.01 by nonpaired Student’s t test. (P) ** P < 0.01 by nonpaired Student’s t test compared with month 0 body weight. (B) * P ≤ 0.05 by two-way analysis of variance (ANOVA) followed by post hoc Bonferroni tests.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Concentration Assay, Saline, Immunofluorescence, Western Blot, Expressing, Cell Culture, Two Tailed Test

Journal: Science Advances

Article Title: Macrophage-derived chemokine CCL22 establishes local LN-mediated adaptive thermogenesis and energy expenditure

doi: 10.1126/sciadv.adn5229

Figure Lengend Snippet: Primer sequences for qPCR gene expression analysis.

Article Snippet: The plasma CCL22 levels were measured by a CCL22/MDC PicoKine ELISA kit (EK0447, Boster Bio).

Techniques: Gene Expression