Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet: ( A ) Heatmaps showing the expression of differentially expressed genes with p-value <0.05 of chemokines and chemokine receptors in resting tissular tissue-resident memory T-cells (T RM ) and effector memory T-cells (T EM ) from patients with COPD (n=2 independent samples) in comparison with control subjects (n=2 independent samples) (GEO accession GSE61397). Expression values are expressed as Z-score transformed transcript count. ( B ) Migration experiment design. ( C ) Migration of fibrocytes from patients with COPD in response to CD8 + T cells supernatants from control subjects (Ctl NS ) or COPD patients (COPD). n=6 independent experiments ( D ) CCL2, CXCL2, and CXCL8 levels in CD8 + T cells supernatants from non-smoking control subjects (Ctl NS ) or patients with COPD (COPD) using BioPlex (CCL2, CXCL2) or ELISA (CXCL1, 3, 5, 6, 8). n=4 Ctl NS samples, n=4 COPD samples. *p<0.05, Mann-Whitney test. ( E–F ), Migration of fibrocytes from patients with COPD in response to CD8 + T cells supernatants from control subjects (Ctl NS ) or COPD patients (COPD), in the presence of 200 nM Reparixin (+) or corresponding vehicle (−) ( E ), and in the presence of 1 µg/mL blocking antibody for CXCL8 (+) or control antibody (−) ( F ). n=6 independent experiments *p<0.05, Wilcoxon matched pairs test.

Article Snippet: When indicated, supernatants were pretreated for 30 min at 37 °C with blocking Ab against CXCL8 (clone 6217, BioTechne, 1 µg/mL) or respective control Ab.

Techniques: Expressing, Comparison, Transformation Assay, Migration, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Blocking Assay

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet: Fibrocyte chemotaxis towards CD8 + T cells is mainly due to an increased CXCL8 secretion by CD8 + T cells in COPD lungs, and promotes direct contact between both cell types. This interaction triggers CD8 + T cell proliferation, cytokine production, and cytotoxic activity. The interaction and its consequences might be further increased by a reinforcement of IFN-γ signaling and expression of molecules belonging to the immune synapse, from the fibrocyte side.

Article Snippet: When indicated, supernatants were pretreated for 30 min at 37 °C with blocking Ab against CXCL8 (clone 6217, BioTechne, 1 µg/mL) or respective control Ab.

Techniques: Chemotaxis Assay, Activity Assay, Expressing

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet:

Article Snippet: When indicated, supernatants were pretreated for 30 min at 37 °C with blocking Ab against CXCL8 (clone 6217, BioTechne, 1 µg/mL) or respective control Ab.

Techniques: Blocking Assay, Recombinant

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: a , Western blot analysis of CXCR2 expression in the negative control (NC), positive control (PC) and CM. Top: the NC is mouse embryonic fibroblasts, the PC is a human embryonic kidney cell line transfected with a Cxcr2 expression vector and the CM are cardiomyocytes of neonatal rats. Bottom: the NC is undifferentiated P19CL6 (mouse embryonic carcinoma) cells, the PC is a human embryonic kidney cell line transfected with a Cxcr2 expression vector and the CM are cardiomyocytes of mice. The experiment was repeated independently three times with similar results. b , Relative mRNA expression of HF markers in cardiomyocytes after addition of the medium conditioned by MYC overexpression in CFs, with and without a neutralizing antibody against CXCR2 ( n = 3 each). Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (GFP ( n = 6) versus MYC ( n = 10), Nppa ), P = 0.0027 (MYC versus MYC plus anti-CXCR2 ( n = 5), Nppa ), P < 0.001 (GFP ( n = 7) versus MYC ( n = 10), Nppb ), P = 0.0027 (MYC versus MYC plus anti-CXCR2 ( n = 5), Nppb ), P < 0.001 (GFP ( n = 12) versus MYC ( n = 9), Myh7 ), P = 0.0040 (MYC versus MYC plus anti-CXCR2 ( n = 5), Myh7 ); significance was determined using the two-sided Mann–Whitney test for mean comparisons. c , Relative mRNA expression of HF markers measured by RT-qPCR in the hearts of control, TAC and TAC with neutralizing antibody against CXCR2. Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (control ( n = 10) versus TAC ( n = 7), Nppa ), P = 0.017 (TAC versus TAC plus anti-CXCR2 ( n = 3), Nppa ), P = 0.0020 (control ( n = 10) versus TAC ( n = 4), Nppb ), P = 0.057 (TAC versus TAC plus anti-CXCR2 ( n = 3), Nppb ), P < 0.001 (control ( n = 10) versus TAC ( n = 6), Myh7 ), P = 0.038 (TAC versus TAC plus anti-CXCR2 ( n = 4), Myh7 ); significance was determined using the two-sided Mann–Whitney test for mean comparisons. d , Bar plots showing cardiac function evaluated by echocardiography in TAC mice with (anti-CXCR2) and without neutralizing antibody against CXCR2 (control). Data are shown as mean ± s.d. (TAC0w, n = 8 (control) and 7 (anti-CXCR2); TAC2w, 6 and 6; TAC4w, 6 and 6; TAC8w, 6 and 7; TAC12w, 8 and 7; TAC14w, 5 and 6; TAC16w, 7 and 6). * P < 0.05; P = 0.036 (TAC12w), P = 0.0037 (TAC14w), P = 0.015 (TAC16w); significance was determined using the two-sided Holm–Sidak test for mean comparisons. e , Comparison of cardiomyocyte size between control and TAC mice with and without neutralizing antibody against CXCR2 (anti-CXCR2). Cell size was evaluated using WGA staining. Quantitative analyses of the cell size in each mouse are shown ( n = 100 cells each). Box plots show the median (center line) and first and third quartiles (box edges), while the whiskers go from each quartile to the minimum or maximum. * P < 0.05; P < 0.001 (control versus TAC), P < 0.001 (TAC versus TAC + anti-CXCR2); significance was determined using the two-sided Mann–Whitney test for mean comparisons. f , Comparison of fibrotic areas between control and TAC mice with and without neutralizing antibody against CXCR2. Fibrosis was evaluated in the perivascular and interstitial regions by azan staining. Scale bars, 300 μm. Representative histological data and quantitative analyses of the fibrotic areas in each mouse are shown ( n = 7 each). Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (control versus TAC, perivascular), P = 0.54 (TAC versus TAC plus anti-CXCR2, perivascular), P < 0.001 (control versus TAC, interstitial), P = 0.38 (TAC versus TAC plus anti-CXCR2, interstitial); significance was determined using the two-sided Mann–Whitney test for mean comparisons. g , Bar plots showing cardiac function evaluated by echocardiography in control, and shRNA of CXCR2-injected mice (shCXCR2) after TAC. Data are shown as mean ± s.d. ( n = 7 (control) and 5 (shCXCR2)). * P < 0.05; P = 0.025 (TAC8w); significance was determined using the two-sided Holm–Sidak test for mean comparisons. h , t -SNE plot of single-cell transcriptomes of human hearts (control: n = 1,673; HF: n = 29,496) (colored by cell clusters). Endo, endothelial cell; SM, smooth muscle cell; MC, myeloid cell; LC, lymphoid cell. i , t -SNE unsupervised clustering of fibroblasts in the human heart colored by cell clusters. j , Cell ratio of MYC -expressing CFs (control: n = 2; dilated cardiomyopathy (DCM): n = 4). Box plots show the median (center line) and first and third quartiles (box edges), while the whiskers go from each quartile to the minimum or maximum. k , Immunofluorescence for PDGFRα and smFISH for MYC and CXCL1 in human control hearts and DCM hearts. Scale bars, 20 μm. The experiment was repeated independently three times with similar results. The arrows indicate the colocalization of the MYC, CXCL1 and PDGFRα in the same cells. l , Bar plots showing the effect of CXCL1 on the contractile properties of iPSCM organoids. The plots show the rate of change of tissue contraction kinetics. Each parameter was normalized by the value of the control well ( n = 14 each). Data are shown as mean ± s.d. * P < 0.05; P = 0.047; significance was determined using the two-sided unpaired t -test for mean comparisons.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Western Blot, Expressing, Negative Control, Positive Control, Transfection, Plasmid Preparation, Over Expression, MANN-WHITNEY, Quantitative RT-PCR, Control, Comparison, Staining, shRNA, Injection, Immunofluorescence

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: Related to Fig. . a : Experimental scheme of TAC mice injected with a neutralising antibody against CXCR2. b : Bar plots showing left ventricle chamber size and wall thickness measured by echocardiography in control and anti-CXCR2 mice after TAC. Mean and standard error are shown (TAC0w, n = 8 [control] and 7 [anti-CXCR2]; TAC2w, 6 and 6; TAC4w, 6 and 6; TAC8w, 6 and 7; TAC12w, 8 and 7; TAC14w, 5 and 6; TAC16w, 7 and 6). *, p < 0.05; p = 0.040 (LVDs, TAC16w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. c : Bar plots showing left ventricle chamber size and wall thickness measured by echocardiography in control and shCXCR2 mice after TAC. Mean and standard error are shown (n = 6 [control] and 5 [shCXCR2] for TAC0w, TAC2w, TAC4w, TAC6w, TAC8w, and TAC10w). *, p < 0.05; p = 0.015 (LVDd, TAC10w), p = 0.0020 (LVDs, TAC10w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. d : Bar plots showing cardiac function, left ventricle chamber size, and wall thickness measured by echocardiography in control and shRNA of Cxcx1 injected mice (shCXCL1) after TAC. Mean and standard error are shown (n = 7 [control] and 5 [shCXCL1] for TAC0w, TAC2w, TAC4w, TAC6w, TAC8w, and TAC10w). *, p < 0.05; p = 0.0064 (LVDd, TAC10w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. e : Left panel, Western blot analysis of p-ERK, ERK, and GAPDH expression in cardiomyocytes from neonatal rats treated with CXCL1 alone or in combination with the anti-CXCR2 antibody. Representative data are shown. Right panel, quantification of Western blot optical densities, with p-ERK/ERK ratios displayed. *, p < 0.05; significance was determined using the two-sided Mann-Whitney test for mean comparisons. Source data are provided as a Source Data file.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Injection, Control, shRNA, Western Blot, Expressing, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: Related to Fig. . a : Immunofluorescence for PDGFRα and single-molecule fluorescence in situ hybridization for MYC and CXCL1 in human dilated phase HCM (dHCM) and sarcoidosis hearts. Scale bars, 20 μm. Arrows indicate the colocalization of the MYC, CXCL1, and PDGFRα in the same cells. b : CXCR2 expression data using single-cell RNA-seq from our previous report. Horizontal axis, TNNT2 expression; vertical axis, CXCR2 expression. Black squares, cardiomyocytes from human control hearts; blue triangles, cardiomyocytes from human hearts of dilated cardiomyopathy. c : Western blot analysis of CXCR2 and GAPDH expression in iPSCM. d : Relative mRNA expression of HF markers in iPSCM (n = 3–5 each). Data are shown as mean ± SD. *, p < 0.05; p = 0.029 (Nppa), p = 0.036 (Nppb); significance was determined using the two-sided Mann-Whitney test for mean comparisons. Source data are provided as a Source Data file.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Immunofluorescence, Fluorescence, In Situ Hybridization, Expressing, RNA Sequencing, Control, Western Blot, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: FACS analyses of CD3, CD11b, and Ly6G expression in mice hearts of sham (TAC0w), TAC4w, TAC8w, TAC12w, and TAC 12w with neutralising antibody against CXCR2.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Quercetin alleviates ulcerative colitis through inhibiting CXCL8-CXCR1/2 axis: a network and transcriptome analysis

doi: 10.3389/fphar.2024.1485255

Figure Lengend Snippet: FIGURE 2 Expression changes of CXCL8-CXCR1/2 in UC tissue versus normal colon tissue. (A) Multiplex fluorescent immunohistochemistry was used to measure CXCL8, CXCR1, and CXCR2 protein levels in normal colon tissue (NCT) and ulcerative colitis tissues (UCT). CXCL8 was labeled in green, CXCR1 was labeled in red, CXCR2 was labeled purple, and DAPI was used to label nuclei in blue (magnification ×200); (B–D) Changes in AOD (average optical density) protein expression levels of CXCL8-CXCR1/2 in different intestinal tissues (n = 5). *p < 0.05, **p < 0.01, ***p < 0.001 vs. NCT.

Article Snippet: The samples were then separated by SDS-PAGE, transferred to a PVDFmembrane, blocked with 5% BSA, and incubated with a primary antibody at 4°C for 14 h (CXCL8, Proteintech, diluted 1:1,000; CXCR1, Affinity, diluted 1: 1,000; CXCR2, Proteintech, diluted 1:1,000; IL-17A, BOSTER, diluted 1:1,000; NF-κB, Affinity, diluted 1:1,000; ZO-1, Affinity, diluted 1:2,000; MUC2, Abcam, diluted 1:2,000; Occludin, Affinity, diluted 1:1,000; GAPDH, Servicebio, diluted 1:4,000).

Techniques: Expressing, Multiplex Assay, Immunohistochemistry, Labeling

Journal: Frontiers in Pharmacology

Article Title: Quercetin alleviates ulcerative colitis through inhibiting CXCL8-CXCR1/2 axis: a network and transcriptome analysis

doi: 10.3389/fphar.2024.1485255

Figure Lengend Snippet: FIGURE 3 CXCL8-CXCR1/2 gene expression in the transcriptome analysis. (A) KEGG pathway enrichment reveals the UC’s top 20 significantly enriched pathways group. (B–D) GSEA results compare the UC group to the HC group, using the Normalized Enrichment Score (NES) as a key metric. NES >0 indicates core genes on the left of the peak, while NES <0 indicates core genes on the right. Significance of the Enrichment Score (ES) is assessed using the p-value.

Article Snippet: The samples were then separated by SDS-PAGE, transferred to a PVDFmembrane, blocked with 5% BSA, and incubated with a primary antibody at 4°C for 14 h (CXCL8, Proteintech, diluted 1:1,000; CXCR1, Affinity, diluted 1: 1,000; CXCR2, Proteintech, diluted 1:1,000; IL-17A, BOSTER, diluted 1:1,000; NF-κB, Affinity, diluted 1:1,000; ZO-1, Affinity, diluted 1:2,000; MUC2, Abcam, diluted 1:2,000; Occludin, Affinity, diluted 1:1,000; GAPDH, Servicebio, diluted 1:4,000).

Techniques: Gene Expression

Journal: Frontiers in Pharmacology

Article Title: Quercetin alleviates ulcerative colitis through inhibiting CXCL8-CXCR1/2 axis: a network and transcriptome analysis

doi: 10.3389/fphar.2024.1485255

Figure Lengend Snippet: FIGURE 6 Que regulated the CXCL8-CXCR1/2 axis. (A–D) CXCL8, CXCR1, and CXCR2 expression levels in mice’s colonic tissue (n = 4). (E–H) IHC expression of CXCL8, CXCR1, and CXCR2 in the colon (×100, ×400) (n = 4).*p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The samples were then separated by SDS-PAGE, transferred to a PVDFmembrane, blocked with 5% BSA, and incubated with a primary antibody at 4°C for 14 h (CXCL8, Proteintech, diluted 1:1,000; CXCR1, Affinity, diluted 1: 1,000; CXCR2, Proteintech, diluted 1:1,000; IL-17A, BOSTER, diluted 1:1,000; NF-κB, Affinity, diluted 1:1,000; ZO-1, Affinity, diluted 1:2,000; MUC2, Abcam, diluted 1:2,000; Occludin, Affinity, diluted 1:1,000; GAPDH, Servicebio, diluted 1:4,000).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Quercetin alleviates ulcerative colitis through inhibiting CXCL8-CXCR1/2 axis: a network and transcriptome analysis

doi: 10.3389/fphar.2024.1485255

Figure Lengend Snippet: FIGURE 8 Workflow for analyzing Que’s mechanisms in UC treatment. Network pharmacology, in conjunction with transcriptomic methodologies and experimental validation, demonstrated that quercetin alleviated the symptoms associated with DSS-induced colitis. It attenuates the inflammatory response, promotes the repair of colonic mucosal tissues, and elevates the levels of colonic tight junction proteins in mice with ulcerative colitis. These effects may enhance tight junction integrity and restore intestinal barrier function by modulating the IL-17A signaling pathway while simultaneously inhibiting the CXCL8-CXCR1/2 axis.

Article Snippet: The samples were then separated by SDS-PAGE, transferred to a PVDFmembrane, blocked with 5% BSA, and incubated with a primary antibody at 4°C for 14 h (CXCL8, Proteintech, diluted 1:1,000; CXCR1, Affinity, diluted 1: 1,000; CXCR2, Proteintech, diluted 1:1,000; IL-17A, BOSTER, diluted 1:1,000; NF-κB, Affinity, diluted 1:1,000; ZO-1, Affinity, diluted 1:2,000; MUC2, Abcam, diluted 1:2,000; Occludin, Affinity, diluted 1:1,000; GAPDH, Servicebio, diluted 1:4,000).

Techniques: Biomarker Discovery