Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a The mean fluorescence intensity (MFI) of CXCR4 in peripheral neutrophils from healthy controls (n = 24) and psoriasis patients (n = 54). b Correlation of the CXCR4 MFI on peripheral neutrophils with PASI in psoriasis patients (n = 25). c The proportion of circulating CD15 + CXCR4 hi neutrophils in healthy controls (n = 20) and psoriasis patients (n = 25). d Correlation of the proportions of CXCR4 hi neutrophils with PASI in psoriasis patients. The adjusted R 2 and P -values were plotted in the graph. e Representative immunoblots of total CXCR4 in circulating neutrophils from healthy controls and psoriasis patients. The relative multiple expression was counted. Blots for each antigen were processed in the same experiment in parallel. The result was repeated twice independently with similar results. f Immunofluorescence staining of CD15 (green) and CXCR4 (red) in normal and inflamed psoriatic skin. Scale bar = 50 µm. n = 10 biologically independent samples. g The proportion of CXCR4 lo vs . CXCR4 hi neutrophils in inflamed psoriatic skin (n = 30 fields from 10 patient samples). MFI of CXCR4 on peripheral neutrophils ( h ) and serum protein levels of CXCL12, IL-17A, and MPO ( i ) before and after treatment with anti-IL-17 inhibitor for 12 weeks. n = 5 biologically independent samples. Data are mean ± SD. Analyses: unpaired Student’s t-test in ( a) and ( c) ; The Spearman method in ( b ) and ( d ); Paired Student’s t-test in ( h ) and ( i ). The paired and unpaired Student’s t-test were conducted as two-sided tests. FMO, Fluorescence Minus One; HC, healthy control; MFI, mean fluorescence intensity; Pre, pre-treatment; Pso, psoriasis patients. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Fluorescence, Western Blot, Expressing, Immunofluorescence, Staining, Control

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Representative images and quantification of different nucleus morphology of CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. Scale bar = 5 µm. b Flow cytometry analysis of key immune markers between peripheral CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients (scale: log-transformed MFI). c Assessment of CXCR4 hi neutrophil survival by Annexin V and 7-AAD staining after 24 h of culture. d Measurement of ROS production by DHE fluorescence. e Quantification of NETs in CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. Phagocytosis of pHrodo Green E. coli by neutrophils was evaluated by immunofluorescence staining ( f ) and flow cytometry ( g ). Scale bar = 5 µm. h Degranulation of CXCR4 lo or CXCR4 hi neutrophils as assessed by CD63 expression. ELISA quantification ( i ) and representative immunoblots ( j ) for mature MMP-9 in the supernatant of CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients that cultured for 24 h. GAPDH was analyzed from corresponding neutrophils. Blots for each antigen were processed in the same experiment in parallel. k Relative mRNA expression of pro-inflammatory mediators in CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. Data are mean ± SD (n = 6 biologically independent samples). The immunofluorescence staining was repeated three times independently with similar results. Two-way ANOVA with Tukey’s post hoc test was performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant. HC, healthy control; MFI, mean fluorescence intensity; MMP-9, matrix metalloprotein 9; NETs; neutrophil extracellular traps; neu, neutrophils; Pso, psoriasis patients. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Flow Cytometry, Transformation Assay, Staining, Fluorescence, Immunofluorescence, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Cell Culture, Control

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Volcano plot showing the number of differentially expressed genes (DEGs) between CXCR4 lo and CXCR4 hi neutrophils from healthy controls (n = 7) and psoriasis patients (n = 6). b Heatmap of DEGs between psoriatic CXCR4 lo and CXCR4 hi neutrophils related to neutrophil and immune function. c Enriched GO terms between CXCR4 lo and CXCR4 hi psoriatic neutrophils. d Gene set enrichment analysis showing enrichment of genes involved in glycolysis in psoriatic CXCR4 hi neutrophils. e Flow cytometry of glycolytic markers in CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. f Uptake of glucose (2-NBDG) in CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. g Extracellular lactate production in CXCR4 lo and CXCR4 hi neutrophils from healthy controls and psoriasis patients. Mean ± SD (n = 6 biologically independent samples/group). Two-way ANOVA with Tukey’s post hoc test was performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant. HC, healthy control; MFI, mean fluorescence intensity; Pso, psoriasis patients; RNA-seq, RNA sequencing. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Flow Cytometry, Control, Fluorescence, RNA Sequencing

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Representative images of adherent CXCR4 lo and CXCR4 hi neutrophils co-cultured with HMEC-1 cells. White arrows indicate HMEC-1 cells; red arrows indicate adherent neutrophils. Scale bar = 10 µm. b Relative mRNA expression of adhesion molecules in HMEC-1 cells with indicated treatment. Representative immunoblots ( c ) and qRT-PCR analysis ( d ) of tight junctions in HMEC-1 cells co-cultured with indicated treatment. e Stimulation of HMEC-1 cells with CXCR4 lo and CXCR4 hi neutrophils for 6 h, followed by analysis of leaked fluorescence intensity of FITC-dextran in a Transwell system. Representative immunoblots of HMEC-1 cells pre-incubated with LDHA inhibitor for 30 min following co-culture with psoriatic CXCR4 hi neutrophils ( f ) and qRT-PCR assessment of tight junction genes ( g ). h Analysis of released FITC-dextran in a Transwell system with HMEC-1 cells, pre-incubated with LDHA inhibitor for 30 min, followed by co-culture with psoriatic CXCR4 hi neutrophils. i Confocal images of CD31 (green) and GPR81 (purple) in psoriatic lesions (n = 6) with CD15 + CXCR4 hi neutrophils adjacent to GPR81 + vascular ECs. Scale bar = 50 µm, 20 µm. The result was repeated three times independently with similar results. HMEC-1 cells were transfected with GPR81 siRNA and subjected to indicated treatment, followed by Western blot ( j ), qRT-PCR ( k ), and FITC-dextran leakage ( l ). The immunoblotting samples shown are from the same experiment ( c , f and j ) and blots were processed in parallel. Mean ± SD (n = 6 biologically independent samples/group). Analyses: two-way ANOVA with Tukey’s post hoc test in ( b ), ( d ), ( e ), ( h ) and ( l ); One-way ANOVA with Tukey’s post hoc test in ( g ) and ( k ). One or two-way ANOVA tests were performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant. HC, healthy control; HMEC-1 cells, human microvascular endothelial cells; Pso, psoriasis patients. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Cell Culture, Expressing, Western Blot, Quantitative RT-PCR, Fluorescence, Incubation, Co-Culture Assay, Transfection, Control

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Expression of CXCR4 on neutrophils stimulated with IL-25, CXCL12, and TNF for 2 h was measured by flow cytometry. b Serum level of CXCL12 in healthy controls (n = 24) and psoriasis patients (n = 30) was detected by ELISA. c Expression of CXCR4 on neutrophils with indicated treatment was measured by flow cytometry. d Representative immunofluorescence staining of CD31 (yellow), Vimentin (green), and CXCL12 (purple) in inflamed psoriatic skin (n = 6). Scale bar = 50 µm, 20 µm. e Representative immunofluorescence staining of CD31 (yellow), CXCR4 (green), and CXCL12 (purple) in inflamed psoriatic skin (n = 6). Scale bar = 50 µm, 20 µm. QRT-PCR analysis ( f ) and flow cytometry analysis ( g ) of CXCR4 in healthy neutrophils with indicated treatments. h Representative immunofluorescence co-staining of CXCR4 (red) and LAMP1 (green) in neutrophils. Scale bar = 2 µm. i Examples of low colocalization values in a sample of CXCR4 hi and CXCR4 lo neutrophils of psoriasis patients stained for LCN2 (red) and CD63 (yellow) was visualized by ImageStream analysis. Images are from one representative experiment out of six. The scale bar indicates 7 μm. Isolated neutrophils were used for ( a ), ( c ), ( f ), and ( g ). Mean ± SD (n = 6 biologically independent samples/group). The immunofluorescence staining was repeated three times independently with similar results. Analyses: one-way ANOVA with Tukey’s post hoc test in ( a) and ( c) ; two-way ANOVA with Tukey’s post hoc test in ( f ) and ( g ); Unpaired Student’s t-test in ( b ). The unpaired Student’s t-test was conducted as two-sided tests. One or two-way ANOVA tests were performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant. FMO, Fluorescence Minus One; HC, healthy control; LCN2, lipocalin 2; MFI, mean fluorescence intensity; Pso, psoriasis patients. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Quantitative RT-PCR, Isolation, Fluorescence, Control

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Conceptual network diagram of genes showing the relationship between CREB1 and enrichment pathways, and heatmap of top transcription factors in CXCR4 hi neutrophils. The mRNA expression of CREB1 ( b ) and protein levels of p-CREB1 (s133) ( c ) in CXCR4 lo and CXCR4 hi neutrophils. d Flow cytometry analysis of p-CREB1 (s133) in neutrophils treated with indicated stimulation. e Co-localization of CXCR4 (green) and p-CREB1 (red) in neutrophils from healthy controls and psoriasis patients. Scale bar = 2 µm. Representative staining ( f ) and Western blot ( g ) of CBP (green) and p-CREB1 (red) in neutrophils treated with indicated stimulation for 2 h. h , i Healthy neutrophils were pre-treated with CREB1 inhibitor (KG-501, 300 μM) for 1 h, followed by indicated stimulation for 2 h. Flow cytometry analysis of the expression of membrane molecules ( h ), as well as glycolysis ( i ). j Representative co-staining of DNA (Hoechst), Cit-H3 (citrullinated histone-3, red), and MPO (myeloperoxidase, green) to assess NETs formation in psoriatic neutrophils in vitro after pretreatment with KG-501 (300 μM) for 1 h. Scale bar = 20 µm. k Western blot of dHL-60 cells transfected with CREB1 siRNA and subjected to the indicated treatment. l Depiction of the CREB1 binding site in the CXCR4 promoter for ChIP assay. m The luciferase activities of wild type CXCR4 and CXCR4 with the CREB1-binding site mutant were determined by luciferase reporter gene assays in dHL-60 cells. Isolated neutrophils were used for ( b ), ( d ), ( h ), and ( i ), and whole blood was used for ( c ). Mean ± SD (n = 6 biologically independent samples). The immunofluorescence staining was repeated three times independently with similar results. Blots for each antigen were processed in the same experiment in parallel. Two-way ANOVA with Tukey’s post hoc test ( b, c, h, i , and m ) and one-way ANOVA with Tukey’s post hoc test ( d ) were performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant. HC, healthy control; MFI, mean fluorescence intensity; NETs, neutrophil extracellular traps; Pso, psoriasis patients. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Expressing, Flow Cytometry, Staining, Western Blot, Membrane, In Vitro, Transfection, Binding Assay, Luciferase, Mutagenesis, Isolation, Immunofluorescence, Control, Fluorescence

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Relative mRNA expressions of CXCR4 and CXL12 in mice tissues was evaluated by qRT-PCR. n = 6 mice. b Proportion of Ly6G + CXCR4 hi neutrophils in mice skin was determined by flow cytometry. n = 6 mice. c Serum level of CXCL12 in control and IMQ-treated mice was detected by ELISA. n = 6 mice. d Schematic diagram of mouse experimental protocol. IMQ mice were injected intraperitoneally with an anti-Ly6G antibody every other day and then injected subcutaneously with fresh isolated homologous Ly6G + CXCR4 lo or Ly6G + CXCR4 hi neutrophils daily. e Phenotype and representative H&E staining of IMQ-treated mice in indicated groups on day 5. Images are representative of six individual mouse per group. Control group was topically applied with Vaseline cream. Bar = 200 µm. n = 6 mice. f Epidermal thickness was assessed by H&E. n = 20 vision fields from 6 mice. g Representative immunofluorescence staining of CD31 (red) in inflamed skin. Scale bar = 50 µm. n = 6 mice. h Quantification of the dermal vascular area in the H&E-stained sections. n = 10 vision fields from 6 mice. i Relative mRNA expressions of inflammatory cytokines in mice tissues. n = 6 mice. The immunofluorescence staining was repeated three times independently with similar results. Mean ± SD. Analyses: unpaired Student’s t-test in ( a ), ( b ), and ( c ); One-way ANOVA with Tukey’s post hoc test in ( f ), ( h ), and ( i ). The unpaired Student’s t-test was conducted as two-sided tests. One-way ANOVA test was performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant; IMQ, imiquimod. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Quantitative RT-PCR, Flow Cytometry, Control, Enzyme-linked Immunosorbent Assay, Injection, Isolation, Staining, Cream, Immunofluorescence

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: a Phenotype and H&E staining of IMQ-treated mice in different treatment groups. Images are representative of six individual mice per group. The mice in the control group was topically applied with vaseline cream. Bar = 200 µm. n = 6 mice. b Epidermal thickness as assessed by H&E staining. n = 20 vision fields from 6 mice. c Proportion of Ly6G + CXCR4 hi neutrophils in inflamed skin was determined by flow cytometry. n = 6 mice. d Representative immunofluorescence staining of CD31 (red) in lesional skin of indicated groups. Scale bar = 50 µm. n = 6 mice. e Quantification of the dermal vascular area in the H&E-stained sections. n = 10 vision fields from 6 mice. f Quantification of extracted Evans blue dye in inflamed skin in indicated groups. n = 6 mice. g Relative mRNA expressions of inflammatory cytokines in mice tissues. n = 6 mice. The immunofluorescence staining was repeated three times independently with similar results. Mean ± SD. One-way ANOVA with Tukey’s post hoc test was performed as two-sided analyses and adjusted for multiple comparisons in the statistical analyses. ns, not significant; IMQ, imiquimod. Source data are provided as a Source Data file.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Staining, Control, Cream, Flow Cytometry, Immunofluorescence

Journal: Nature Communications

Article Title: CREB1-driven CXCR4 hi neutrophils promote skin inflammation in mouse models and human patients

doi: 10.1038/s41467-023-41484-3

Figure Lengend Snippet: This study provides insights into the development and role of CXCR4 hi neutrophils in skin inflammation. Compared to CXCR4 lo neutrophils, CXCR4 hi neutrophils exhibit increased capacity for NETs formation, phagocytosis, and degranulation, as well as higher expression of pro-inflammatory mediators, supported by activated glycolysis. CXCR4 expression on neutrophils is induced by CXCL12, IL-25, and TNF, which requires de novo mRNA and protein synthesis and intracellular protein transport, but is not stored in neutrophil granules and regulated by degranulation. CREB1 is activated by phosphorylation on Ser-133 upon stimulation, enabling interaction with its coactivator protein CBP to initiate transcription of CREB-responsive genes, thereby contributing to CXCR4 expression and CXCR4 hi neutrophils in skin inflammation. Our data further emphasize the crucial role of CXCR4 hi neutrophils in promoting vascular remodeling through lactate and MMP-9 release (B), facilitating immune cell infiltration into tissues, and increasing inflammatory responses (A). Moreover, we identify the CXCR4/CXCL12 axis as a potential therapeutic target for inflammatory skin diseases, with a focus on CREB1, CXCR4/CXCL12, or the formation of NETs. CBP, CREB-binding protein; EC, endothelial cell; MMP9, matrix metallopeptidase 9; NETs; neutrophil extracellular traps; PAD4 i, PAD4 inhibitor. Created with BioRender.com.

Article Snippet: CXCR4 hi neutrophils were also obtained by positive selection from total neutrophils using mouse CXCR4 MicroBeads (130-118-682, Miltenyi Biotec Inc.) according to the manufacturer’s protocol, as depicted below.

Techniques: Expressing, Phospho-proteomics, Binding Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Representative images of MC38-OVA (left) and tumour growth (right) in C57BL/6J mice ( n = 5). b . Representative images of CT26 tumours (left) and tumour growth (right) in BALB/c mice ( n = 5). c . Schematic diagram illustrating the working principle of ATS-GNP adipocytolysis. ATS-GNP, containing a CaCO 3 core, is receptor-mediated for endocytosis. Upon reaching the acidic environment, ATS-GNP releases CO 2 gas, disrupting the adipose cell membrane. d . Tumour growth of MC38 tumours in C57BL/6J, BALB/c-Nude (left), and NSG (right) mice ( n = 5). e . Representative images of E0771 tumours at day 16 of the experiment in C57BL/6J mice following removal of PAT ( n = 5). f . Tumour weights of E0771 tumours at day 16 in C57BL/6J mice with the removal of PAT ( n = 5). g . Representative flow cytometry plots of CD45 + cells gated on live cells in MC38-OVA tumours. h . Violin plots showing the expression of CXCR7 across all cell types in CRC patients, analysed using the Kruskal-Wallis test. i . Editing strategy for constructing Cxcl12 fl/fl cKO mouse. j . Experimental design for constructing Control and Cxcl12 fl/fl cKO mice bearing MC38-OVA tumours near PAT. k . Western blot analysis for verifying the knockout efficiency of Cxcl12 in PAT of Control mice and Cxcl12 fl/fl cKO mice. l . RT-qPCR analysis for verifying the Cxcl12 knockout efficiency in liver, spleen, uterus, and tumour tissues from Control mice and Cxcl12 fl/fl cKO mice ( n = 6). Data represent ≥ 3 independent experiments. P -values were calculated using two-way ANOVA with Tukey’s correction for multiple comparisons ( a -right, b -right, and d ), two-way ANOVA with Bonferroni’s correction for multiple comparisons ( f ), or a two-sided, unpaired Student’s t -test ( l ). Graphs display mean ± SD ( a , b , d , f , l ). Panels created with BioRender : c , i and j , Huaiqiang, J. https://biorender.com/cy7rgjm (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Membrane, Flow Cytometry, Expressing, Control, Western Blot, Knock-Out, Quantitative RT-PCR

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , Cell–cell communication analysis based on ligand–receptor interactions (top six) between stromal cells and lymphocytes in tVAT (left) and a comparison between tVAT, dVAT and tumour (right). b , Marked CXCL12–CXCR4 interactions among CD8 + T cells, CD4 + T cells, B cells, plasma cells and stromal cell populations in tVAT, dVAT and tumour. The width of the lines represents the probability of communication. c , Violin plots showing the expression of CXCL12 (top) and CXCR4 (bottom) across all cell types in patients with CRC. d , Violin plots comparing the expression of CXCL12 in dVAT versus tVAT (top) and tumour versus tVAT (bottom) in patients with CRC, analysed using a two-sided Wilcoxon test. e , Experimental design for the PAT C57BL/6J mouse model treated with IgG or anti-CXCL12 antibody (left), and representative MC38 tumour images at day 16 of the experiment (right) ( n = 5). f , Tumour growth (left) and tumour weights (right) of MC38 tumours at day 16 of the experiment in C57BL/6J mice ( n = 5). g , Representative MC38 tumour images (left) and tumour growth (right) of experiments in control and Cxcl12 fl/fl cKO mice ( n = 6). h , Tumour weights of MC38 tumours in control and Cxcl12 fl/fl cKO mice at day 16 of the experiment ( n = 6). i , Flow cytometry analysis of the infiltration of various CXCR4 + immune cells in MC38 tumours in Control and Cxcl12 fl/fl cKO mice ( n = 6). j , Schematic diagram of the chemotaxis assay using T cells as ‘sensors’ and conditional medium as a ‘sink’ (left), and the aggregated trajectories of control or CXCL12-induced T cells migrating for 1 h (right). k , Quantitative analysis of CXCR4 + CD45.1 + T cells in MC38 tumours with and without removal of PAT or contralateral inguinal adipose tissue (control) by flow cytometry ( n = 5). Data represent ≥3 independent experiments. Statistical significance was assessed by a two-sided permutation test ( a ), two-sided unpaired Student’s t -test ( f right, h and i ), one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons ( k right) or two-way ANOVA ( f left and g right). Graphs display mean ± s.d. ( f – i , k ). Panels created with BioRender : e and k , Huaiqiang, J. https://biorender.com/ovq2e39 (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Comparison, Clinical Proteomics, Expressing, Control, Flow Cytometry, Chemotaxis Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . UMAP plot of VAT-associated stromal cells in tVAT and dVAT from CRC patients. The 8 clusters, labelled by inferred cell types, are denoted by colour. b . Dot plot showing RNA expression of marker genes used to define pAC, adCAF, and APC subclusters. Circle size represents the log-normalized P -value, while colour intensity indicates the log-transformed mean expression of marker genes. c . Sorting strategy for isolating adCAFs from mouse PAT by flow cytometry. d . RNA sequencing analysis of adCAF markers in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 4). Statistical significance was assessed using a two-sided, unpaired Student’s t -test. e . RNA sequencing analysis of adipogenesis markers in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 4). Statistical significance was assessed using a two-sided, unpaired Student’s t -test. f . Representative images of the morphology of sorted adCAF-enriched stromal cells and non-adCAF stromal cells. Scale bar = 500 μm. g . Cross-tissue interactions analysis based on ligand-receptor pairs between adCAFs and tumour cells (left) and a comparison between tVAT and dVAT (right). A two-sided permutation test was used to determine the significance of pathways. h . Western blot analysis of CXCL12 protein expression in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 3). The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median) for ( d - e ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: RNA Expression, Marker, Transformation Assay, Expressing, Flow Cytometry, RNA Sequencing, Comparison, Western Blot, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , UMAP of all stromal cells in tVAT, dVAT, tumour and normal from patients with CRC, with ten clusters labelled by inferred cell types. Major lineages included ASCs, pACs, CAFs, pericytes (PCs) and mesothelial cells (Mesos). b , Heatmap displaying the distribution of eight stromal cell subtypes across different tissue types. c , UMAP of eight subsets of VAT-associated stromal cells in tVAT and dVAT from patients with CRC, including ASCs, pACs and adCAFs. d , Beeswarm plot showing the distribution and abundance of VAT-associated stromal cell types in Nhoods between tVAT and dVAT. e , Stack plot displaying the abundance of the eight VAT-associated stromal cell subsets in dVAT and tVAT. f , Heatmap showing the RNA expression of various marker genes in VAT-associated stromal cell types, including ASC/pAC markers, CAF markers, cytokines and stromal markers. g , Representative multiplex immunofluorescence images showing the presence of adCAFs in tVAT samples from patients with CRC. Scale bar, 10 μm. DAPI, 4,6-diamidino-2-phenylindole. h , Relative expression levels of multiple cytokines and protumoural factors in adCAF-enriched stromal cells ( n = 4) and non-adCAF stromal cells ( n = 4) derived from the PAT of mice xenograft models. The data are presented as a box-and-whisker graph (bounds of box show first to third quartile, bottom and top line show minimum to maximum and the central line shows the median). i , Cell–cell communication analysis based on ligand–receptor pairs (top six) between adCAFs and lymphocytes in tVAT (left) and a comparison between tVAT and dVAT (right). j , RT–qPCR (left) and ELISA (right) detecting the RNA expression and protein secretion of CXCL12 in sorted adCAF-enriched stromal cells and non-adCAF stromal cells from PAT of mice. Data represent ≥3 independent experiments. All data are shown as mean ± s.d. and statistical significance was assessed by a two-sided, unpaired Wilcoxon test ( h ), two-sided permutation test ( i ) and Student’s t -test ( j ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: RNA Expression, Marker, Multiplex Assay, Immunofluorescence, Expressing, Derivative Assay, Whisker Assay, Comparison, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . UMAP of all cells in tVAT and dVAT from 5 CRC patients, with 16 clusters labelled by inferred cell types. Major lineages included various immunocytes, VAT-associated stromal cells, adipocytes (ACs), endothelial cells (ECs), pericytes (PCs) and mesothelial cells (Mesos). b . Heatmap of representative marker genes across all cell populations. c . UMAP of 6 subsets of ACs in tVAT and dVAT from 5 CRC patients. d . Beeswarm plot showing the distribution and abundance of ACs in Nhoods between tVAT and dVAT. e . Box plot to compare the abundance of ACs in dVAT ( n = 5) and tVAT ( n = 5). P -values were calculated using a two-side Wilcoxon test. f . Cell-cell communication analysis based on ligand-receptor interactions (top 3) between CD8 + T cells and adipocytes, and between CD8 + T cells and VAT-associated stromal cells in tVAT. A two-sided permutation test was used to determine the significance of pathways. Only significant ligand-receptor pairs ( P < 0.05) are shown. Dot size encodes the interaction score, and colour scale indicates the communication probability. g . Violin plots showing the expression of CXCL12, VAT-associated markers, and CAF-associated markers in adipocytes and VAT-associated stromal cells from CRC patients. h . UMAP of 8 subsets of VAT-associated stromal cells in tVAT and dVAT from 5 CRC patients, including APCs, pACs and adipocyte-derived cancer-associated fibroasts (adCAFs). The 8 clusters, labelled by inferred cell types, are denoted by colour. i . Beeswarm plot showing the distribution and abundance of VAT-associated stromal cell types in Nhoods between tVAT ( n = 5) and dVAT ( n = 5). j . Box plot to compare the abundance of VAT-associated stromal cells in dVAT and tVAT. P -values were calculated using a two-side Wilcoxon test. k . Heatmap showing the RNA expression patterns of representative marker genes across all cell populations, including CAF markers, stromal markers, APC/pAC markers, white/brown/beige fat markers, and cytokines and growth markers. The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median) for ( e , j ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Marker, Expressing, Derivative Assay, RNA Expression, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , Experimental design for constructing Control and Mdk DTR cKO mice bearing MC38 tumours near PAT, followed by αPD-1 therapy. b , c , Representative MC38 tumour images ( b ), tumour weights ( c left) and tumour growth ( c right) in control and Mdk DTR cKO mice treated with IgG or anti-PD-1 ( n = 6). d , Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells, CXCR4 + immunocytes, CXCR4 + T cell, CXCR4 + CD4 + T cell, CXCR4 + CD8 + T cell and CXCR4 + tumour-specific T cell in MC38-OVA tumours from the four treatment groups ( n = 6). e , f , Representative MC38 tumour images ( e ) and tumour weights ( f ) of the experiment in mice treated with anti-CXCL12 and/or anti-PD-1 ( n = 5). g , Representative MRI image of CRC tumour and corresponding tVAT area region of CR and non-CR patients pre- and post-immuno-chemoradiotherapy. The yellow area represents the tVAT area, whereas the red area denotes the tumour region. Note that the mass visible in the intestinal lumen (top right) is faecal material. h , Pre-treatment tVAT area difference based on 3D Slicer between CR ( n = 30) and non-CR ( n = 37) patients. The data are presented as a box-and-whisker graph (bounds of box show first to third quartile, bottom and top line show minimum to maximum and the central line shows the median). i , ROC plot of response predicting ability of pre-treatment PAT area in immuno-chemoradiotherapy of proficient mismatch repair patients with CRC, compared with conventional indexes, including CPS, TPS, CEA and CA199 ( n = 67) with optimal cutoff. j , Comparison of pCR ratio in tVAT high and low group according to the optimal cutoff. k , Graphical abstract depicting how tumours reshape the stromal environment in tVAT and how tVAT competes for immunocytes from the tumour to promote immune escape. Data represent ≥3 independent experiments. Statistical significance was assessed using a two-sided, unpaired Student’s t -test ( d , h ), one-way ANOVA with Tukey’s correction for multiple comparisons ( c left, f ) or two-way ANOVA with Tukey’s correction for multiple comparisons ( c right). Graphs display mean ± s.d. ( c , d , f , h ). Panels created with BioRender : a and k , Huaiqiang, J. https://biorender.com/e5jwcye (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Control, Flow Cytometry, Whisker Assay, Comparison

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Editing strategy for constructing Mdk DTR mouse. b . Flow cytometry representative plots and bar graphs for verifying the elimination efficiency of adCAFs (FAP + PDGFRB + MDK + stromal cells) in PAT of Control mice and Mdk DTR cKO mice. c . Experimental design for combination therapy with αCXCL12 and αPD-1 in mice bearing MC38-OVA tumours near PAT. d . MC38 tumour weights of the experiment in mice treated with αCXCL12 and/or αPD-1 ( n = 6). e . Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells in MC38-OVA tumours from the 4 treatment groups ( n = 6). f . Flow cytometry analysis of the infiltration of CXCR4 + immunocytes, including CXCR4 + T cell, CXCR4 + CD4 + T cell and CXCR4 + CD8 + T cell in MC38-OVA tumours from the 4 treatment groups ( n = 6). g . Boxplot of difference of tVAT area in CR and non-CR patients with T3 or T4 stage separately. The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median). h . ROC plot of prediction ability of tVAT area in patients with T3 or T4 stage separately. Data represent ≥ 3 independent experiments. P -values were calculated using a two-sided, unpaired Student’s t -test ( b , e - g ) and two-way ANOVA with Tukey’s correction for multiple comparisons ( d ). Graphs display mean ± SD ( b , d , e - g ). Panels created with BioRender : a and c , Huaiqiang, J. https://biorender.com/33r2gmm (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Flow Cytometry, Control, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Experimental design for combination therapy with AMD3100 and αPD-1 in mice bearing MC38-OVA tumours near subcutaneous PAT. b . Representative MC38 tumour images of the mice treated with AMD3100 and/or αPD-1 ( n = 5). c . Representative MC38 tumour weights of the mice treated with AMD3100 and/or αPD-1 ( n = 5). d . Representative MC38 tumour growth of the mice treated with AMD3100 and/or αPD-1 ( n = 5). e . Representative MC38 tumour images in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). f . Representative MC38 tumour weights in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). g . Representative MC38 Bioluminescence Images in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). h . Representative MC38 tumour images in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). i . Representative MC38 tumour weights in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). j . Representative MC38 Bioluminescence Images in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). k . Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells in MC38-OVA tumours from 4 treatment groups ( n = 5). l . Flow cytometry analysis of the infiltration of CXCR4 + immunocytes, including CXCR4 + T cell, CXCR4 + CD4 + T cell and CXCR4 + CD8 + T cell in MC38-OVA tumours from 4 treatment groups ( n = 5). Data represent ≥ 3 independent experiments. P -values were calculated using a two-sided, unpaired Student’s t -test ( c , f , i , k , l ) and two-way ANOVA with Tukey’s correction for multiple comparisons ( d ). Graphs display mean ± SD ( c - d , f , i , k - l ). Panel created with BioRender : a , Huaiqiang, J. https://biorender.com/wbcolts (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Flow Cytometry

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Expression of CXCR4 and CXCL12 and their correlations to the cell proliferation and angiogenesis in mycosis fungoides

doi: 10.5114/pdia.2015.48034

Figure Lengend Snippet: Expression of CXCL12 and CXCR4 in mycosis fungoides (MF) and healthy skin (IRS – immunoreactive score)

Article Snippet: Following commercial primary antibodies against studied proteins were used for the immunohistochemical staining: CXCR4: Monoclonal Anti-human CXCR4 (fusin) Antibody; Clone: 44716; Ig class: IgG2B; Catalog Number: MAB 172; R&D SYSTEMS; dilution 1: 120; CXCL12: Monoclonal Anti-human/mouse CXCL12/SDF-1 Antibody; Clone: 79018; Ig class; mouse IgG1; Catalog Number: MAB 350; R&D SYSTEMS USA; dilution 1: 60.

Techniques: Expressing, Staining

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Expression of CXCR4 and CXCL12 and their correlations to the cell proliferation and angiogenesis in mycosis fungoides

doi: 10.5114/pdia.2015.48034

Figure Lengend Snippet: Correlations between the expression of CXCL12, CXCR4 and proliferation markers (ρ – Spearman correlation coefficient) in MF

Article Snippet: Following commercial primary antibodies against studied proteins were used for the immunohistochemical staining: CXCR4: Monoclonal Anti-human CXCR4 (fusin) Antibody; Clone: 44716; Ig class: IgG2B; Catalog Number: MAB 172; R&D SYSTEMS; dilution 1: 120; CXCL12: Monoclonal Anti-human/mouse CXCL12/SDF-1 Antibody; Clone: 79018; Ig class; mouse IgG1; Catalog Number: MAB 350; R&D SYSTEMS USA; dilution 1: 60.

Techniques: Expressing

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Expression of CXCR4 and CXCL12 and their correlations to the cell proliferation and angiogenesis in mycosis fungoides

doi: 10.5114/pdia.2015.48034

Figure Lengend Snippet: Comparison of the expression of Ki67, CD34, AgNORs, CXCL12, CXCR4 with the disease course and prognosis

Article Snippet: Following commercial primary antibodies against studied proteins were used for the immunohistochemical staining: CXCR4: Monoclonal Anti-human CXCR4 (fusin) Antibody; Clone: 44716; Ig class: IgG2B; Catalog Number: MAB 172; R&D SYSTEMS; dilution 1: 120; CXCL12: Monoclonal Anti-human/mouse CXCL12/SDF-1 Antibody; Clone: 79018; Ig class; mouse IgG1; Catalog Number: MAB 350; R&D SYSTEMS USA; dilution 1: 60.

Techniques: Comparison, Expressing

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CAFs induce sorafenib resistance in HCC cells by secreting CXCL12. a The results of immunofluorescence showed that the expression of CXCL12 in CAFs in HCC tissues was significantly higher than that in paracancerous tissues (left). Statistical plot of fluorescence intensity of fibroblasts expressing α-SMA and CXCL12 in HCC tissues and paracancerous tissues (right). b ELISA showed that CAFs secreted higher level of CXCL12 than NFs. c , d Colony forming assays detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with the cellular supernatant of CAFs and NFs, sorafenib, and AMD3100. e , f Flow cytometry apoptosis assay detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with the cellular supernatant of CAFs and NFs, sorafenib, and AMD3100. g , h Western blotting was performed to detect the expression of β-actin, and Cleaved Caspase-3 in HCC cells (HepG2 and Huh7), which were treated with the cellular supernatant of CAFs, sorafenib, and AMD3100. The data presented mean ± SEM. * p < 0.01; ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: Immunofluorescence, Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Apoptosis Assay, Western Blot

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CXCL12 induces sorafenib resistance in HCC cells by up-regulating the expression of FOLR1. a We found two datasets of cancer cells treated with CXCL12 protein (GSE15893 and GSE40017) in the GEO database. We took the intersection of the differentially expressed genes between these two datasets and combined them with the reported drug-resistant genes of HCC to obtain two genes. FOLR1 was the most significantly upregulated drug resistance-related gene upon CXCL12 treatment. b The qPCR was performed to detect the level of FOLR1 in Huh7 and HepG2, which treated with CXCL12 protein and AMD3100. c Western blotting was performed to detect the expression of β-actin and CXCR4 in CXCR4 knockdown HCC cells (Huh7 and HepG2). d , e Western blotting was performed to detect the expression of β-actin, CXCR4, FOLR1, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, CXCL12 protein, and AMD3100. f , g Western blotting was performed to detect the expression of β-actin, CXCR4, FOLR1, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, AMD3100, the supernatant of CAFs, and NFs. h , i Colony forming assay detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with sorafenib, CXCL12 protein, the supernatant of CAFs, and AMD3100. j , k Western blotting was performed to detect the expression of β-actin, FOLR1, CXCR4, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, anti-CXCR4, CXCL12 protein, and the supernatant of CAFs. The data presented mean ± SEM. ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: Expressing, Western Blot, Knockdown

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CAFs enhance sorafenib resistance of HCC cells through CXCL12 in vivo. a Representative images of tumors in mice of CAFs + AMD3100 group, CAFs group, and NFs group after different treatments. b The tumor volume in different treatment groups. c The tumor proliferation trend in different treatment groups. d Pathological validation of tumors under a microscope (40X), after H&E staining and Immunohistochemistry in tumor tissues. The immunohistochemistry staining to detect the expression of Cleaved Caspase-3 in different treatment groups from the tumor tissues of mice. e The expression level of Cleaved Caspase-3 in different treatment groups from the tumors of mice. The data presented mean ± SEM. ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: In Vivo, Biomarker Discovery, Microscopy, Staining, Immunohistochemistry, Expressing