recombinant cxcl13  (R&D Systems)

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

Journal: Frontiers in Immunology

Article Title: CXCR5 engineered human and murine Tregs for targeted suppression in secondary and tertiary lymphoid organs

doi: 10.3389/fimmu.2025.1513009

Figure Lengend Snippet: FVIII TRuCε CXCR5 Tregs display improved in vitro and in vivo persistence. (A) In vitro migration of FVIII CAR, FVIII CAR CXCR5, and FVIII TRuCε transduced Tregs through a transwell in response to either serum free media, CXCL12 or CXCL13 gradients. Number of migrated mScarlet + cells at the bottom of the transwell are quantified by flow cytometry following 6hrs of incubation. (B) Kinetics of in vivo migration of adoptively transferred FVIII TRuCε or FVIII TRuCε CXCR5 T conv cells to the spleen on days 1, 2, 4, and 7 following adoptive transfer. Mice received i.v. injections of recombinant FVIII on days 0, 3, and 6. Frequencies of mScarlet + cells per total CD4 + T cells are quantified by flow cytometry. (C) Number of mScarlet + FVIII TRuCε or FVIII TRuCε CXCR5 Tregs per 10 7 CD4 + T cells are quantified from spleens and (D) inguinal lymph nodes (ILN) on day 7 post adoptive transfer. Data represents mean±SEM, ****p<0.0001, ∗p < 0.05, ∗∗p < 0.01 using 2-way ANOVA with Tukey’s multiple comparisons analysis for (A) , 2-way ANOVA with Sidak’s multiple comparisons analysis for (B) , unpaired t test for (C, D) .

Article Snippet: Lower chambers contained 600 μL serum-free medium with 1μg/mL of either recombinant mouse CXCL12 or CXCL13 (PeproTech, Rocky Hill, NJ).

Techniques: In Vitro, In Vivo, Migration, Flow Cytometry, Incubation, Adoptive Transfer Assay, Recombinant