Journal: Nature Cancer

Article Title: Reprogramming of stroma-derived chemokine networks drives the loss of tissue organization in nodal B cell lymphoma

doi: 10.1038/s43018-026-01136-z

Figure Lengend Snippet: a , Left, UMAP of LN stromal cells by entity. Right, percentages of rFRCs and rBECs (rLN n = 5, FL n = 6, DLBCL n = 8 patients). b , Differentially expressed genes between rLN-derived ( n = 2,363 cells) and DLBCL-derived ( n = 2,983 cells) FRCs (adjusted P < 0.05, log(fold change) > 0.5). c , Homeostatic (top) and inflammatory (bottom) chemokine expression in bulk data . d , Pearson correlation of CXCL13 expression and CIBERSORTx-derived FDC fractions. e , CXCL13 plasma protein levels in FL ( n = 18 patients) and DLBCL ( n = 22 patients). Vertical lines indicate the mean per entity. f , Exemplary rLN and DLBCL mIF images, representative of n = 4 patients per entity. Scale bar, 50 μm. Dashed circles: CD21+ regions. g , mIF-derived CXCL13 and CXCR5 signals averaged across four adjacent pairs of CD21 + follicular and CD21 − extrafollicular regions per sample. h , i , Spatial transcriptomics plots of FL-LN ( h ) and DLBCL-LN ( i ) cores colored by cell type and CXCL13 – CXCR5 ligand–receptor (L–R) score. j , mIF-derived enrichment of CXCL13 + cells per cell type in DLBCL versus rLN/FL samples. Asterisks indicate P < 0.01; exact P values are provided in the source data. k , CXCL13 expression in CD8 + T cells ( n = 21,268 cells) . l , Percentage of CXCR5 + cells within CD3 − fractions measured by flow cytometry (rLN n = 7, FL n = 24, DLBCL n = 18 patients). m , Migrated rLN- and DLBCL-derived B cells in the Transwell assay (mean ± s.d., n = 3 patients per condition). For c and d : tonsil n = 10, FL 1/2/3A n = 145, FL 3B n = 48, DLBCL n = 430 patients. For f , g and j : rLN n = 4, FL n = 5, DLBCL n = 4 patients. P values in a , c , e , g and l : two-sided Wilcoxon rank-sum test. P value in m : two-sided unpaired Welch’s t test. P values in j : two-sided Fisher’s exact test. P values in c , g and j were adjusted using the Benjamini–Hochberg method. Box plots: center line, median; box, interquartile range; whiskers, 1.5× the interquartile range; points, data values. FC, fold change; T tox EM, effector memory cytotoxic T cells; hr, human recombinant.

Article Snippet: Sorted cells were seeded in triplicate into the top chambers of 96-well HTS Transwell plates with 8-μm pores (Corning), with the bottom chambers containing either 1 μg ml −1 human recombinant CXCL13 (cat. no. 801-CX-025, R&D Systems) or PBS as a control.

Techniques: Derivative Assay, Expressing, Clinical Proteomics, Spatial Transcriptomics, Flow Cytometry, Transwell Assay, Recombinant

Journal: Nature Cancer

Article Title: Reprogramming of stroma-derived chemokine networks drives the loss of tissue organization in nodal B cell lymphoma

doi: 10.1038/s43018-026-01136-z

Figure Lengend Snippet: ( A ) Heatmap showing estimated cell type fractions as determined using CIBERSORTx (CSx) within a large bulk transcriptomics dataset . ( B ) Box plots depicting CSx fractions of exemplary cell types across disease entities. Boxes indicate the median (center line), interquartile range (bounds) and 1.5x interquartile range (whiskers). Data points represent patient samples. P-values were calculated using two-sided unpaired Welch’s t-test and adjusted according to Benjamini-Hochberg. ( C ) Dot plot showing Spearman correlation coefficients of chemokine expression and cell type fractions. Dot sizes represent -log10 FDR-adjusted p-value of correlation tests. ( D ) Scatter plots color-coded by disease entity correlating CXCL12 (left panel) and CCL21 (right panel) expression with CSx-derived FRC factions. P-values were calculated using Pearson correlation. ( E , F ) Bulk RNA-seq datasets , stratified by FDC abundance into high and low samples using maximally selected rank statistics based on CSx fractions, represented in a Kaplan-Meier curve showing overall survival (left panels; log-rank test) and scatter plot of FDC fractions and CXCL13 expression (right panels; Pearson correlation). T PR =T prol . For panels A – D : tonsil n = 10; FL 1/2/3 A n = 145; FL 3B n = 48; DLBCL n = 430 patients. For panel E : FDC-high n = 61; FDC-low n = 44 patients. For panel F : FDC-high n = 302; FDC-low n = 68 patients.

Article Snippet: Sorted cells were seeded in triplicate into the top chambers of 96-well HTS Transwell plates with 8-μm pores (Corning), with the bottom chambers containing either 1 μg ml −1 human recombinant CXCL13 (cat. no. 801-CX-025, R&D Systems) or PBS as a control.

Techniques: Transcriptomics, Expressing, Derivative Assay, RNA Sequencing

Journal: Nature Cancer

Article Title: Reprogramming of stroma-derived chemokine networks drives the loss of tissue organization in nodal B cell lymphoma

doi: 10.1038/s43018-026-01136-z

Figure Lengend Snippet: ( A ) Representative triangle-thresholded mIF images showing CD21 (top panels) as well as CXCL13 (bottom panels) signal in rLN (n = 4), FL (n = 5) and DLBCL (n = 4) patient samples. Scale bar indicates 50μm. ( B ) UMAP representation of spatial transcriptomics data colored by cell type (color code depicted in panel C ). ( C ) Heatmap showing scaled expression of key marker gene expression as well chemokines. ( D ) Stacked bar plot of FDC/FRC/rFRC fractions (top panel) and violin plot of CXCL13 expression in these subsets (bottom panel). ( E ) Spatial transcriptomics plots of FL (left panels) and DLBCL (right panels) tissue cores colored by cell type (left panels) alongside magnified views of chemokine expression. ( F ) Heatmap showing scaled expression of key T cell markers across CD8 + T cell populations as measured using mIF. ( G ) Box plot showing per-sample percentage of CXCL13 + cells (as determined using Otsu thresholding) among PD1 + CD8 + effector memory T cells in the mIF dataset. Boxes indicate the median (center line), interquartile range (bounds) and 1.5x interquartile range (whiskers). Data points represent patient samples. P-values were calculated using a two-sided unpaired Welch’s t-test. ( H ) Spearman correlation coefficients of key T cell markers across all CD8 + T cell populations (n = 21,268 cells). ( I ) Volcano plot showing differentially expressed genes comparing PD1 + CD8 + effector memory T cell populations (T TOX EM-II n = 2,018; T TOX EM-III n = 9,208 cells). Labels indicate cluster-identifying genes as well as CXCL13 . Abbreviations: T TOX EM = effector memory cytotoxic T cells. For panels A , F , G : rLN n = 4; FL n = 5; DLBCL n = 4 patients. For panels B – E : FL n = 1; DLBCL n = 1 patient.

Article Snippet: Sorted cells were seeded in triplicate into the top chambers of 96-well HTS Transwell plates with 8-μm pores (Corning), with the bottom chambers containing either 1 μg ml −1 human recombinant CXCL13 (cat. no. 801-CX-025, R&D Systems) or PBS as a control.

Techniques: Spatial Transcriptomics, Expressing, Marker, Gene Expression

Journal: Nature Cancer

Article Title: Reprogramming of stroma-derived chemokine networks drives the loss of tissue organization in nodal B cell lymphoma

doi: 10.1038/s43018-026-01136-z

Figure Lengend Snippet: a , UMAP representation of a microarray dataset with homeostatic ( CXCL12 , CXCL13 , CCL19 , CCL21 ) and inflammatory ( CXCL9 , CXCL10 , CXCL11 ) chemokine expression values used as features for dimensionality reduction. Left, UMAP displaying pie charts (within each dot) that represent the k = 20 nearest neighbors, colored according to disease entity. Right, the same UMAP colored according to the mean expression of homeostatic (top) and inflammatory (bottom) chemokines. b , Bulk RNA-seq dataset stratified according to homeostatic chemokine expression into high ( n = 519 patients) and low ( n = 99 patients) groups using maximally selected rank statistics, shown as a dot plot (top) and a Kaplan–Meier curve of overall survival (bottom). The P value was calculated using the log-rank test. c , Forest plot summarizing log 10 -transformed hazard ratios (center), 95% confidence intervals (error bars) and Wald-derived P values estimated from univariate Cox proportional hazards models assessing the association between homeostatic chemokine expression and overall survival across five individual DLBCL bulk datasets , – . d , Same UMAP as in a , colored by CIBERSORTx-derived FDC fractions. e , Bulk RNA-seq dataset stratified by FDC abundance into high ( n = 255 patients) and low ( n = 364 patients) groups using maximally selected rank statistics based on CIBERSORTx fractions, shown as a Kaplan–Meier curve of overall survival (left; log-rank test) and a scatter plot of log-transformed FDC fractions and CXCL13 expression (right; Pearson correlation). f , Forest plot summarizing log 10 -transformed hazard ratios (center), 95% confidence intervals (error bars) and Wald-derived P values estimated from Cox proportional hazards models assessing the association between FDC fraction and overall survival across five individual DLBCL bulk datasets , – . For panels a and d : tonsil n = 10, FL 1/2/3A n = 145, FL 3B n = 48 and DLBCL n = 430 patients. OS, overall survival; HR, hazard ratio.

Article Snippet: Sorted cells were seeded in triplicate into the top chambers of 96-well HTS Transwell plates with 8-μm pores (Corning), with the bottom chambers containing either 1 μg ml −1 human recombinant CXCL13 (cat. no. 801-CX-025, R&D Systems) or PBS as a control.

Techniques: Microarray, Expressing, RNA Sequencing, Transformation Assay, Derivative Assay

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Construction and characterization of the rRABV expressing CXCL13. (A) Schematic diagram for the construction of LBNSE, LBNSE-GM-CSF, and LBNSE-CXCL13. The mouse CXCL13 gene was cloned and inserted into the RABV genome in place of the deleted long noncoding region, and rRABVs were rescued according to the method described in Materials and Methods. (B) A multistep growth curve was generated in BSR cells. Cells were infected with LBNSE, LBNSE-GM-CSF, or LBNSE-CXCL13 at a multiplicity of infection (MOI) of 0.01 FFU and incubated at 37°C. Viruses were harvested at 1, 2, 3, 4, and 5 dpi, and viral titers were determined. All titrations were carried out in quadruplicate, and the data are presented as the means ± standard deviations (SD). (C) Production of CXCL13 in BSR cells. Cells were infected with different viruses at MOIs of 0.001, 0.01, 0.1, and 1. After incubation at 37°C for 24 h, the culture supernatants were collected, and the CXCL13 concentrations produced by the indicated rRABVs were determined with a commercial ELISA kit. (D) Chemotactic effects of cultured medium from BSR cells infected with rRABVs at an MOI of 1 on mouse splenocytes. Splenocytes (5 × 105) were applied to the upper wells of chemotaxis chambers. Two and 4 h later, the cells migrating to the bottom chamber were counted. (E and F) BALB/c mice were inoculated via i.m. injection of 1 × 106 FFU of rRABVs. The muscles from the hind legs of mice (n = 3) were harvested at 3 and 6 dpi. Total RNA was extracted, and viral genomic RNA (vRNA) (E) and CXCL13 mRNA and CXCR5 mRNA (F) were analyzed via qRT-PCR. All the data are expressed as the means ± SD. Asterisks indicate significant differences between the indicated experimental groups.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Expressing, Clone Assay, Generated, Infection, Incubation, Produced, Enzyme-linked Immunosorbent Assay, Cell Culture, Chemotaxis Assay, Injection, Muscles, Quantitative RT-PCR

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Recruitment of Tfh cells by CXCL13. BALB/c mice (n = 3) were infected via i.m. injection of 1 × 106 FFU of different rRABVs, and the spleens, draining LNs, and blood were harvested at 7 and 14 dpi. Single-cell suspensions were prepared, stained with antibodies against Tfh cells and Tfh cell activation markers, and analyzed via flow cytometry. (A and B) Representative gating strategies for the detection of Tfh cells (A) and representative flow cytometric plots of Tfh cells (B) are shown. (C to E) The results of a detailed analysis for activated Tfh cells (CD4+ CXCR5+ PD-1+) at 7 and 14 dpi are presented for the spleen (C), the draining LNs (D), and the blood (E). Data are expressed as the means ± SEM (n = 3). Asterisks indicate significant differences between the indicated experimental groups.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Infection, Injection, Staining, Activation Assay, Flow Cytometry

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Recruitment of GC B cells by CXCL13. BALB/c mice (n = 3) were infected via i.m. injection of 1 × 106 FFU of different rRABVs, and the spleens and draining LNs were harvested at 7 and 14 dpi. Single-cell suspensions were prepared, stained with antibodies against GC B cells and GC B cell activation markers, and analyzed via flow cytometry. (A and B) Representative gating strategies for the detection of GC B cells (A) and representative flow cytometric plots of GC B cells (B) are shown. (C and D) The results of a detailed analysis for activated GC B cells (B220+ CD95+ GL7+) at 7 and 14 dpi are presented for the spleen (C) and the draining LNs (D). Data are presented as the means ± SEM (n = 3). Asterisks indicate significant differences between the indicated experimental groups.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Infection, Injection, Staining, Activation Assay, Flow Cytometry

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Expression of CXCL13 facilitates the formation of GCs. BALB/c mice (n = 3) were infected via i.m. injection of 1 × 106 FFU of different rRABVs, and the draining LNs were collected at 7 and 14 dpi. Then, the draining LNs were excised, and tissue sections were prepared and stained for germinal centers (GL7, red; B220, blue; and IgG, green). Scale bars, 500 μm or 100 μm (rightmost column only). (A) Representative results are shown. (B) The numbers of GCs formed at 7 and 14 dpi were calculated. All the data are expressed as the means ± SEM (n = 3). Asterisks indicate significant differences between the indicated experimental groups; ns, not significant.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Expressing, Infection, Injection, Staining

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Plasma CXCL13 levels correlate with GC activity and VNA titers in mice. BALB/c mice were infected via i.m. injection of 1 × 106 FFU of DMEM (n = 5), LBNSE (n = 9), LBNSE-GM-CSF (n = 9), or LBNSE-CXCL13 (n = 9), and then draining (inguinal) and nondraining (cervical) LNs were collected at 7 dpi. Single-cell suspensions were prepared, stained with antibodies against GC B cells and GC B cell activation markers, and analyzed via flow cytometry. (A and B) Representative gating strategies for the detection of GC B cells (A) and representative flow cytometric plots of plasma B cells (B) are shown. (C) The results of a detailed analysis of activated GC B cells (CD20+ Ki-67+ Bcl-6+) at 7 dpi are presented for the draining and nondraining LNs. (D) Serum samples were harvested at 7 dpi, and RABV VNA titers were measured via FAVN tests as described in Materials and Methods. (E) The concentration of plasma CXCL13 was determined using a commercial ELISA kit. (F) Correlations of GC B cell activity in the draining LNs and RABV VNA titers in mice 7 days after immunization were determined. (G) Correlations of plasma CXCL13 concentrations and RABV VNA titers in mice 7 days after immunization were determined. (H) Correlations of GC B cell activity in the draining LNs and plasma CXCL13 concentrations in mice 7 days after immunization were determined. All the data are expressed as the means ± SEM (n = 3). Asterisks indicate significant differences between the indicated experimental groups.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Clinical Proteomics, Activity Assay, Infection, Injection, Staining, Activation Assay, Flow Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: Expression of CXCL13 promotes an increase in the quantity of plasma cells. BALB/c mice (n = 3) were infected via i.m. injection of 1 × 106 FFU of different rRABVs, and the bone marrow samples were harvested at 7 and 14 dpi. Single-cell suspensions were prepared, stained with antibodies against plasma B cells and plasma B cell activation markers, and analyzed via flow cytometry. (A and B) Representative gating strategies for the detection of plasma B cells (A) and representative flow cytometric plots of plasma B cells (B) are shown. (C) The results of a detailed analysis of activated plasma B cells (B220low CD138+) at 7 and 14 dpi are presented for the bone marrow samples. All the data are expressed as the means ± SEM (n = 3). Asterisks indicate significant differences between the indicated experimental groups.

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Expressing, Clinical Proteomics, Infection, Injection, Staining, Activation Assay, Flow Cytometry

Journal: Journal of Virology

Article Title: A Novel Rabies Vaccine Expressing CXCL13 Enhances Humoral Immunity by Recruiting both T Follicular Helper and Germinal Center B Cells

doi: 10.1128/JVI.01956-16

Figure Lengend Snippet: qRT-PCR primers used in this study

Article Snippet: Recombinant mouse CXCL13 proteins were purchased from R&D Systems (USA).

Techniques: Sequencing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: CXCR4 inhibition in human pancreatic and colorectal cancers induces an integrated immune response

doi: 10.1073/pnas.2013644117

Figure Lengend Snippet: The effect of CXCL12-stimulated CXCR4 on chemokine receptor-mediated migration of human immune cells. ( Left ) The coexpression of CXCR4 with ( A ) CXCR1, ( B ) CXCR3, ( C ) CXCR5, ( D ) CXCR6, and ( E ) CCR2 on human immune cell lines was evaluated by flow cytometry after staining with antibodies specific for the relevant chemokine receptors. Gray peaks indicate isotype controls. ( Center ) The effect of CXCL12-stimulation of CXCR4 on the chemotactic responses of A CXCR1-coexpressing Jurkat T lymphoblastoid cells to CXCL8, ( B ) CXCR3-coexpressing HSB2DP T lymphoblastoid cells to CXCL10, ( C ) CXCR5-coexpressing Raji B lymphoblastoid cells to CXCL13, ( D ) CXCR6-coexpressing Jurkat T lymphoblastoid cells to CXCL16, and ( E ) CCR2-coexpressing Molm13 monocytoid cells to CCL2 was assessed by including CXCL12 in the upper chamber (blue) and the other chemokines in the lower chamber (red) in the Boyden two-chamber assay. ( Right ) The chemotaxis assays were performed with the five cell lines when the placement of the chemokines in the Boyden chambers was reversed. Bar diagrams display mean and SEM (n = 3-4). Statistical analysis by Student’s t test: *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The concentration of each chemokine used is as following: CXCL8 (R&D Systems, 208-IL), 20 ng/mL; CXCL10 (R&D Systems, 266-IP), 1,000 ng/mL; CXCL13 (R&D Systems, 801-CX), 1,000 ng/mL; CXCL16 (R&D Systems, 976-CX), 50 ng/mL; CCL2 (R&D Systems, 279-MC), 200 ng/mL

Techniques: Migration, Flow Cytometry, Staining, Boyden Chamber Assay, Chemotaxis Assay

Journal: Journal of Advanced Research

Article Title: Adiponectin deficiency prevents chronic colitis-associated colonic fibrosis via inhibiting CXCL13 production

doi: 10.1016/j.jare.2024.12.032

Figure Lengend Snippet: Blockage of CXCL13 reduced the severity and colonic fibrosis of DSS-induced chronic colitis in mice. ( A ) Schematic diagram of the experimental procedure. ( B ) The time-dependent curve of body weight change. ( C ) The time-dependent curve of disease activity index (DAI). ( D ) Representative colon morphology and length. ( E ) Representative histological analysis images and pathological score. ( F ) Representative image of Masson trichromatic staining of colon tissue. ( G ) The mRNA expressions of Tgf‐β, MMP-9, Col1a1, Col3a1 and Cxcr5 in the colon tissues of DSS-induced chronic colitis mice. ( H ) The protein expressions of α-SMA, MMP-9 and CXCL13 in the colon tissues of DSS-induced chronic colitis mice. Data represent the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The recombinant lentivirus harboring Cxcl13 shRNA (LV- Cxcl13 shRNA) and NC shRNA (LV-NC) was generated through packaging with the YSH-LV001 vector by Ubigene Biosciences Co., Ltd (Guangzhou, China).

Techniques: Activity Assay, Staining

Journal: Journal of Advanced Research

Article Title: Adiponectin deficiency prevents chronic colitis-associated colonic fibrosis via inhibiting CXCL13 production

doi: 10.1016/j.jare.2024.12.032

Figure Lengend Snippet: APN deficiency prevented CXCL13 production in the colon of mice with chronic colitis. ( A ) Differential genes by RNA-sequence analysis were shown by a volcano map. ( B ) KEGG pathway enrichment analysis were shown with P < 0.05. ( C ) Differential genes related with cytokine-cytokine receptor interaction were shown in a heat map. ( D ) The interaction of differential genes involved in cytokine-cytokine receptor interaction was displayed by PPI network. ( E ) The gene expression of Cxcl13 and its receptor Cxcr5 in the colon tissues of chronic colitis mice was analyzed by RT-qPCR. ( F and G ) Protein expression of CXCL13 in the colon tissues of chronic colitis mice was detected by western blot and ELISA assays, respectively. Data represent the means ± SEM. * P < 0.05 and ** P < 0.01.

Article Snippet: The recombinant lentivirus harboring Cxcl13 shRNA (LV- Cxcl13 shRNA) and NC shRNA (LV-NC) was generated through packaging with the YSH-LV001 vector by Ubigene Biosciences Co., Ltd (Guangzhou, China).

Techniques: Sequencing, Gene Expression, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: Adiponectin deficiency prevents chronic colitis-associated colonic fibrosis via inhibiting CXCL13 production

doi: 10.1016/j.jare.2024.12.032

Figure Lengend Snippet: APN deficiency attenuated the CXCL13-expressing M2 macrophages in the mice with chronic colitis. ( A ) CD206-positive cells infiltrated in the colon tissues of DSS-induced chronic colitis from WT and APN -/- mice. ( B and C ) The accumulation of F4/80 + CD206 + cells in F4/80 + macrophages and F4/80 + CD206 + CXCL13 + cells in F4/80 + CD206 + macrophages within the colonic lamina propria (cLPs) of DSS-induced chronic colitis in WT and APN -/- mice. ( D ) The mRNA expressions of CXCL13 and M2 macrophage markers CD206 , Arg-1 and Fizz1 in F4/80 + cells isolated from colonic lamina propria (cLPs) of DSS-induced chronic colitis from WT and APN-KO mice. ( E ) The protein expressions of p-Akt in in F4/80 + cells isolated from colonic lamina propria (cLPs) of DSS-induced chronic colitis from WT and APN-KO mice. Data represent the means ± SEM. ** P < 0.01 and *** P < 0.001.

Article Snippet: The recombinant lentivirus harboring Cxcl13 shRNA (LV- Cxcl13 shRNA) and NC shRNA (LV-NC) was generated through packaging with the YSH-LV001 vector by Ubigene Biosciences Co., Ltd (Guangzhou, China).

Techniques: Expressing, Isolation

Journal: Journal of Advanced Research

Article Title: Adiponectin deficiency prevents chronic colitis-associated colonic fibrosis via inhibiting CXCL13 production

doi: 10.1016/j.jare.2024.12.032

Figure Lengend Snippet: Exogenous CXCL13 promoted the fibrogenesis in mouse L929 and human CCD-18Co fibroblasts. (A) The mRNA expressions of fibrogenesis-related genes α-SMA, Tgf‐β, MMP-9, Col1a1 and Col3a1 in L929 fibroblasts. ( B ) The fluorescence intensity of COL1A1 in L929 fibroblasts with immunofluorescent staining (× 200, COL1A1 (green) and DAPI (blue)). ( C ) The protein expressions of α-SMA and COL1A1 in L929 fibroblasts. (D) The mRNA expressions of fibrogenesis-related genes α-sma, tgf‐β, and col1a1 in CCD-18Co fibroblasts. ( E ) The protein expressions of α-SMA and COL1A1 in CCD-18Co fibroblasts. ( F ) The protein expressions of p-Akt, p-p38, p-ERK, and p-JNK. ( G ) The mRNA expressions of CXCL13 specific receptor, Cxcr5 . ( H ) Inhibitors targeting Akt (MK-2206 dihydrochloride, 1 μM, MCE), p38 (SB233580, 10 μM, MCE), ERK (U0126, 10 μM, MCE), and JNK (SP600125, 10 μM, MCE) demonstrated a significant reversal of CXCL13-induced upregulation of fibrosis-promoting genes Col1a1 and Col3a1 . ( I ) Blocking CXCR5 with siRNA reversed the up-regulation of CXCL13-induced fibrogenesis-related genes Col1a1 and Col3a1 . Data represent the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The recombinant lentivirus harboring Cxcl13 shRNA (LV- Cxcl13 shRNA) and NC shRNA (LV-NC) was generated through packaging with the YSH-LV001 vector by Ubigene Biosciences Co., Ltd (Guangzhou, China).

Techniques: Fluorescence, Staining, Blocking Assay

Journal: Journal of Advanced Research

Article Title: Adiponectin deficiency prevents chronic colitis-associated colonic fibrosis via inhibiting CXCL13 production

doi: 10.1016/j.jare.2024.12.032

Figure Lengend Snippet: CXCL13 was up-regulated in colon tissues of UC patients and correlated positively with APN expression and colonic fibrosis. (A) The mRNA expression level of CXCL13 was notably upregulated in the sample from UC colonic inflamed mucosa (dataset GSE38713 , GSE9452 and GSE11223 ). Student's t -test was used. (B) Representative images of CXCL13 expression in colon tissue samples from UC patients (IHC method). ( C ) Pearson’s correlation between CXCL13 expression and the colonoscopy score of UC patients. ( D ) Pearson’s correlation between CXCL13 expression and the expressions of APN, COL1A1 and COL3A1 in colon tissue samples from UC patients, respectively. All results were presented as the mean ± SEM. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: The recombinant lentivirus harboring Cxcl13 shRNA (LV- Cxcl13 shRNA) and NC shRNA (LV-NC) was generated through packaging with the YSH-LV001 vector by Ubigene Biosciences Co., Ltd (Guangzhou, China).

Techniques: Expressing