ccl19 antibodies  (Bioss)

Journal: Nature Communications

Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea

doi: 10.1038/s41467-024-52946-7

Figure Lengend Snippet: a UMAP plots showing subclusters and sample conditions of fibroblasts. b Bar graph showing the percentage of subpopulations of fibroblasts. c Feature plot showing the expression of the C3 in fibroblasts. d The percentage of C3 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). e Immunohistochemistry of C3 and DCN in skin samples. Below panels are magnified images of boxed areas. White arrows indicate C3 negative fibroblasts. Yellow arrows indicate C3 positive (C3 + ) fibroblasts. Scale bar, 50 μm. f Quantification of the percentage of C3 + fibroblasts in different groups ( n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in e ). g The top-ranked enriched KEGG pathways in C3 + fibroblasts revealed by GSEA. NES stands for enrichment scores. Two-sided permutation test without multiple comparison adjustments was used for GSEA analysis. h Heatmap showing the expression levels of multiple inflammatory factors in C3 + fibroblasts in rosacea and healthy skins. i The percentage of CCL19 + /CXCL1 + /CXCL2 + /CXCL12 + and C3 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). j Feature plots showing the expression of CCL19, CXCL1, CXCL2, CXCL12 in all cell types. Data are presented as mean ± SEM and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( f ), and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( d , i ).

Article Snippet: The following primary antibodies were used: Goat anti-PDGFRA antibody (1:100, AF1062, R&D systems), Rabbit anti-PTGDS antibody (1:200, 10004344, Cayman), Rabbit anti-PDGFRA (1:500, ab203491, abcam), Goat anti-CCL19 (1:100, AF880, R&D systems), Rabbit anti-CD4 (1:500, ab183685, abcam), Rat anti-CCR7 (1:100, MAB3477, R&D systems), Rat anti-IFNγ (1:100, 14-7311-81, Thermo), Rabbit anti-MBP (1:1500, 78896, Cell signaling), Sheep anti-CD74 (1:100, AF7478, R&D systems), Rat anti-F4/80 (1:100, 14-4801-81, Thermo), Goat anti-CXCL10 antibody (1:100, AF466, R&D systems), Rabbit anti-MLC2 (1:400, 15354-1-AP, Proteintech), Mouse anti-α-SMA (1:5000, ab7817, abcam).

Techniques: Expressing, Immunohistochemistry, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea

doi: 10.1038/s41467-024-52946-7

Figure Lengend Snippet: a The total interaction strength in rosacea and healthy skins analyzed by CellChat. b – d Bubble diagrams showing the incoming/outgoing interaction strength of each cell type in HS ( b ), non-lesional ( c ) and lesional ( d ) skin of rosacea. e KEGG categories of genes enriched in each cell type from the lesional skin of rosacea patients. Two-sided permutation test without multiple comparison adjustments was used for GSEA analysis. f DEGs of fibroblasts (compared to other cells) in the lesional skin of rosacea patients. g Violin plot showing the expression of CCL19 in fibroblasts. h The percentage of CCL19 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). i Multiplex immunohistochemistry of CCL19, DCN, CCR7 and CD4 in skin samples of HS ( n = 5), NS ( n = 5), and lesional skin of ETR ( n = 5), PPR ( n = 5) and PhR ( n = 5). The presented images are representative of each group. Below panels are magnified images of boxed areas. White arrows indicate CD4 + CCR7 + T cells. Yellow arrows indicate CCL19 + DCN + fibroblasts. Scale bar, 50 μm. j Feature plots showing the expression of PTGDS in all cells (left) and fibroblasts (right). k Violin plot showing the expression of PTGDS in fibroblasts. l Immunohistochemistry of PTGDS and Vimentin in skin samples. White arrows indicate Vimentin + PTGDS − fibroblasts. Yellow arrows indicate Vimentin + PTGDS + fibroblasts. Scale bar, 50 μm. m Quantification of the percentage of PTGDS + fibroblasts ( n = 6/6/6/6/6 samples for HS/NS/ETR/PPR/PhR group used in l ). n Levels of PGD 2 in skin samples, detected by ELISA ( n = 11/14/12/11 samples for HS/ ETR/PPR/PhR group). o Immunohistochemistry of PTGDR and α-SMA in skin samples of HS, normal skin of rosacea patients (NS), and lesional skin of ETR, PPR and PhR. Scale bar, 50 μm. Data are presented as mean ± SEM and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( g , k , m ), and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( h , n ).

Article Snippet: The following primary antibodies were used: Goat anti-PDGFRA antibody (1:100, AF1062, R&D systems), Rabbit anti-PTGDS antibody (1:200, 10004344, Cayman), Rabbit anti-PDGFRA (1:500, ab203491, abcam), Goat anti-CCL19 (1:100, AF880, R&D systems), Rabbit anti-CD4 (1:500, ab183685, abcam), Rat anti-CCR7 (1:100, MAB3477, R&D systems), Rat anti-IFNγ (1:100, 14-7311-81, Thermo), Rabbit anti-MBP (1:1500, 78896, Cell signaling), Sheep anti-CD74 (1:100, AF7478, R&D systems), Rat anti-F4/80 (1:100, 14-4801-81, Thermo), Goat anti-CXCL10 antibody (1:100, AF466, R&D systems), Rabbit anti-MLC2 (1:400, 15354-1-AP, Proteintech), Mouse anti-α-SMA (1:5000, ab7817, abcam).

Techniques: Comparison, Expressing, Multiplex Assay, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: JCI Insight

Article Title: CXCL9, CXCL10, and CCL19 synergistically recruit T lymphocytes to skin in lichen planus

doi: 10.1172/jci.insight.179899

Figure Lengend Snippet: ( A ) UMAP depicting subclustering of epidermal skin cells. ( B ) Volcano plots of differential gene expression from basal keratinocyte subpopulations from lichen planus (LP) lesional versus nonlesional skin. Expression of CXCL9 , CXCL10 , and CCL19 is labeled on the plots. A P value of less than 0.01 (Wilcox’s test) and a 2-fold expression change was used for significance. ( C ) UMAP depicting subclustering of fibroblasts. ( D ) Volcano plots of differential gene expression from fibroblast subpopulations from LP lesional versus nonlesional skin. Expression of CXCL9 , CXCL10 , and CCL19 is labeled on the plots. A P value of less than 0.01 (Wilcox’s test) and a 2-fold expression change was used for significance. ( E ) Representative immunofluorescence images depicting localization of CXCL9 (green), CCL19, (white), and DAPI (blue) in LP skin ( n = 5 patient samples). The white dotted line depicts the epidermal-dermal junction. Scale bars: 100 μm. ( F ) scRNA-Seq data from publicly available single-cell datasets for vitiligo, psoriasis, and atopic dermatitis were used to analyze skin cells for their expression of chemokines. Dot size corresponds to percentages of cells expressing chemokine, while color corresponds to level of gene expression.

Article Snippet: For immunofluorescence microscopy, the following antibodies were used: CD3 (MCA1477, Bio-Rad), CD4 + (ab183685, Abcam), CXCL13 (AF801, R&D Biosystems), CXCL9 (ab9720, Abcam), CCL19 (PA5-109488, Thermo Fisher Scientific), and CCL21 (AF366, R&D Biosystems).

Techniques: Gene Expression, Expressing, Labeling, Immunofluorescence

Journal: JCI Insight

Article Title: CXCL9, CXCL10, and CCL19 synergistically recruit T lymphocytes to skin in lichen planus

doi: 10.1172/jci.insight.179899

Figure Lengend Snippet: ( A ) Global analysis of ligand-receptor pathways. Arrows highlight relevant signaling. ( B ) Analysis of cell-to-cell interactions between epidermal keratinocytes cells and immune cells in lichen planus (LP) lesional skin. Dot color illustrates communication probability, and dot size illustrates P value. ( C ) Analysis of cell-to-cell interactions between dermal fibroblasts and immune cells in lesional (salmon color) and nonlesional (blue color) LP skin. Dot color illustrates communication probability, and dot size illustrates P value. ( D ) Schematic of migration assay. ( E – H ) Left: migration index (no. of migrated cells in response to cytokine/no. of migrated cells in response to control) for CD4 + and CD8 + T cells in response to different conditions of CXCL9, CXCL10, and CCL19 ( n = 12). Right: Combined treatment with or without CCR7 blocking antibodies ( n = 9). Data are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, 1-way ANOVA was used for migration assays, and 2-tailed paired Student’s t test was used for CCR7 antibody analysis.

Article Snippet: For immunofluorescence microscopy, the following antibodies were used: CD3 (MCA1477, Bio-Rad), CD4 + (ab183685, Abcam), CXCL13 (AF801, R&D Biosystems), CXCL9 (ab9720, Abcam), CCL19 (PA5-109488, Thermo Fisher Scientific), and CCL21 (AF366, R&D Biosystems).

Techniques: Migration, Control, Blocking Assay

Journal: JCI Insight

Article Title: CXCL9, CXCL10, and CCL19 synergistically recruit T lymphocytes to skin in lichen planus

doi: 10.1172/jci.insight.179899

Figure Lengend Snippet: ( A ) Volcano plots of differential gene expression from exhausted (Texh) and CD8 + proliferating (CD8 + Pro) T cell populations from lesional versus nonlesional lichen planus (LP) skin. Expression of CXCL13 , CTLA4 , GZMB , and GNLY is labeled. ( B ) Representative immunofluorescence images of LP skin depicting localization of CD4 (green), CXCL13 (white), CD3 (red), and DAPI (blue) ( n = 5 patient samples). Scale bars: 100 μm. ( C ) Dot plot demonstrating levels and percentages of cells expressing CXCR3 and CXCR5 . ( D ) Analysis of cell-to-cell interactions between immune cells in lesional (salmon color) and nonlesional (blue color) LP skin. ( E and F ) Left: Migration index (no. of migrated cells in response to cytokine/no. of migrated cells in response to control) for CD4 + ( E ) and CD8 + T cells ( F ) in response to different conditions of CXCL13 and CCL19 ( n = 9). Right: Combined treatment with or without CCR7 blocking antibodies ( n = 7). Data are shown as the mean ± SEM. * P < 0.05, 1-way ANOVA was used for migration assays, and 2-tailed paired Student’s t test was used for CCR7 antibody analysis.

Article Snippet: For immunofluorescence microscopy, the following antibodies were used: CD3 (MCA1477, Bio-Rad), CD4 + (ab183685, Abcam), CXCL13 (AF801, R&D Biosystems), CXCL9 (ab9720, Abcam), CCL19 (PA5-109488, Thermo Fisher Scientific), and CCL21 (AF366, R&D Biosystems).

Techniques: Gene Expression, Expressing, Labeling, Immunofluorescence, Migration, Control, Blocking Assay

Journal: Nature Communications

Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea

doi: 10.1038/s41467-024-52946-7

Figure Lengend Snippet: a UMAP plots showing subclusters and sample conditions of fibroblasts. b Bar graph showing the percentage of subpopulations of fibroblasts. c Feature plot showing the expression of the C3 in fibroblasts. d The percentage of C3 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). e Immunohistochemistry of C3 and DCN in skin samples. Below panels are magnified images of boxed areas. White arrows indicate C3 negative fibroblasts. Yellow arrows indicate C3 positive (C3 + ) fibroblasts. Scale bar, 50 μm. f Quantification of the percentage of C3 + fibroblasts in different groups ( n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in e ). g The top-ranked enriched KEGG pathways in C3 + fibroblasts revealed by GSEA. NES stands for enrichment scores. Two-sided permutation test without multiple comparison adjustments was used for GSEA analysis. h Heatmap showing the expression levels of multiple inflammatory factors in C3 + fibroblasts in rosacea and healthy skins. i The percentage of CCL19 + /CXCL1 + /CXCL2 + /CXCL12 + and C3 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). j Feature plots showing the expression of CCL19, CXCL1, CXCL2, CXCL12 in all cell types. Data are presented as mean ± SEM and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( f ), and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( d , i ).

Article Snippet: The following primary antibodies were used: Rabbit anti-CD31 (1:100, 77699, Cell Signaling), Mouse anti-KRT14 (1:1000, ab7800, abcam), Rabbit anti-KRT14 (1:2000, ab181595, abcam), Rabbit anti-CD74 (1:500, 77274, Cell signaling), Rabbit anti-CD74 (1:2000, ab289885, abcam), Rabbit anti-CLDN4 (1:2000, ab210796, abcam), Rabbit anti-complement C3 (1:400, HPA003563, Atlas Antibodies), Rabbit anti-DCN (1:2000, ab277636, abcam), Rabbit anti-MBP (1:1500, 78896, Cell signaling), Rabbit anti-CD69 (1:500, ab233396, abcam), Rabbit anti-CD103 (1:2000, ab224202, abcam), Rabbit anti-p-MLC2 (1:200, 3671, Cell signaling), Mouse anti-α-SMA (1:5000, ab7817, abcam), Mouse anti-CCL19 (1:100, MAB361, R&D systems), Mouse anti-CD4 (1:200, 14-2444-80, Thermo), Mouse anti-CCR7 (1:100, MAB197, R&D systems), Rabbit anti-PTGDS (1:200, HPA004938, Atlas Antibodies), Rabbit anti-Vimentin (1:400, 5741, Cell signaling), Rabbit anti-PTGDR (1:200, HPA049668, Atlas Antibodies).

Techniques: Expressing, Immunohistochemistry, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea

doi: 10.1038/s41467-024-52946-7

Figure Lengend Snippet: a The total interaction strength in rosacea and healthy skins analyzed by CellChat. b – d Bubble diagrams showing the incoming/outgoing interaction strength of each cell type in HS ( b ), non-lesional ( c ) and lesional ( d ) skin of rosacea. e KEGG categories of genes enriched in each cell type from the lesional skin of rosacea patients. Two-sided permutation test without multiple comparison adjustments was used for GSEA analysis. f DEGs of fibroblasts (compared to other cells) in the lesional skin of rosacea patients. g Violin plot showing the expression of CCL19 in fibroblasts. h The percentage of CCL19 + fibroblasts in total fibroblasts ( n = 3 samples for each group from scRNA-seq datasets). i Multiplex immunohistochemistry of CCL19, DCN, CCR7 and CD4 in skin samples of HS ( n = 5), NS ( n = 5), and lesional skin of ETR ( n = 5), PPR ( n = 5) and PhR ( n = 5). The presented images are representative of each group. Below panels are magnified images of boxed areas. White arrows indicate CD4 + CCR7 + T cells. Yellow arrows indicate CCL19 + DCN + fibroblasts. Scale bar, 50 μm. j Feature plots showing the expression of PTGDS in all cells (left) and fibroblasts (right). k Violin plot showing the expression of PTGDS in fibroblasts. l Immunohistochemistry of PTGDS and Vimentin in skin samples. White arrows indicate Vimentin + PTGDS − fibroblasts. Yellow arrows indicate Vimentin + PTGDS + fibroblasts. Scale bar, 50 μm. m Quantification of the percentage of PTGDS + fibroblasts ( n = 6/6/6/6/6 samples for HS/NS/ETR/PPR/PhR group used in l ). n Levels of PGD 2 in skin samples, detected by ELISA ( n = 11/14/12/11 samples for HS/ ETR/PPR/PhR group). o Immunohistochemistry of PTGDR and α-SMA in skin samples of HS, normal skin of rosacea patients (NS), and lesional skin of ETR, PPR and PhR. Scale bar, 50 μm. Data are presented as mean ± SEM and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( g , k , m ), and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( h , n ).

Article Snippet: The following primary antibodies were used: Rabbit anti-CD31 (1:100, 77699, Cell Signaling), Mouse anti-KRT14 (1:1000, ab7800, abcam), Rabbit anti-KRT14 (1:2000, ab181595, abcam), Rabbit anti-CD74 (1:500, 77274, Cell signaling), Rabbit anti-CD74 (1:2000, ab289885, abcam), Rabbit anti-CLDN4 (1:2000, ab210796, abcam), Rabbit anti-complement C3 (1:400, HPA003563, Atlas Antibodies), Rabbit anti-DCN (1:2000, ab277636, abcam), Rabbit anti-MBP (1:1500, 78896, Cell signaling), Rabbit anti-CD69 (1:500, ab233396, abcam), Rabbit anti-CD103 (1:2000, ab224202, abcam), Rabbit anti-p-MLC2 (1:200, 3671, Cell signaling), Mouse anti-α-SMA (1:5000, ab7817, abcam), Mouse anti-CCL19 (1:100, MAB361, R&D systems), Mouse anti-CD4 (1:200, 14-2444-80, Thermo), Mouse anti-CCR7 (1:100, MAB197, R&D systems), Rabbit anti-PTGDS (1:200, HPA004938, Atlas Antibodies), Rabbit anti-Vimentin (1:400, 5741, Cell signaling), Rabbit anti-PTGDR (1:200, HPA049668, Atlas Antibodies).

Techniques: Comparison, Expressing, Multiplex Assay, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Two Tailed Test