Journal: eLife

Article Title: A non-bactericidal cathelicidin provides prophylactic efficacy against bacterial infection by driving phagocyte influx

doi: 10.7554/eLife.72849

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Mouse CXCL3 ELISA Kit , Rockland , Cat#:KOA0825 , .

Techniques: Flow Cytometry, cDNA Library Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Staining, Blocking Assay, In Vivo, Control

Journal: Journal of Inflammation Research

Article Title: Protective Effects of Qingre Sanjie Jiaonang on Pulmonary Fibrosis: A Pilot Study

doi: 10.2147/JIR.S479432

Figure Lengend Snippet: QRSJ alleviated the release of inflammatory factors during the progression of PF. ( A ) Immunofluorescence staining was performed, with imaging for NE (red), and staining with DAPI for nucleus (blue). Scale bar, 50 μm or 20 μm (Zoom). ( B ) Quantitative analysis of the mean fluorescence intensity of NE (n = 3). ( C ) The effect of QRSJ on the levels of NE in serum in different groups using ELISA (n = 5). ( D-F ) The effect of QRSJ on the levels of CXCL15, IL-1β and TNF-α in BALF in different groups using ELISA (n = 3). ( G ) The effect of QRSJ on the levels of IL-6 in serum in different groups using ELISA (n = 5). ( H ) The effect of QRSJ on the levels of CXCL15 in serum in different groups using ELISA (n = 4). ( I ) The effect of QRSJ on the levels of TNF-α in serum in different groups using ELISA (n = 5). The data is presented as mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 by one-way ANOVA. ns represents no significant difference.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) Kits for tumor necrosis factor (TNF)-α (Cat # E-EL-M3063), IL-6 (Cat # E-EL-M0044), CXCL15 (mouse homologue of human IL-8, Cat # E-EL-M0269), HMGB1 (Cat # E-EL-M0676), IL-1β (Cat # E-EL-M0037) and NE (Cat # E-EL-M3025) were purchased from Elabscience (Wuhan, China).

Techniques: Immunofluorescence, Staining, Imaging, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: METTL3 promotes an immunosuppressive microenvironment in bladder cancer via m6A-dependent CXCL5/CCL5 regulation

doi: 10.1136/jitc-2024-011108

Figure Lengend Snippet: METTL3 regulates the expression and secretion of CXCL5 and CCL5 in bladder cancer. (A) Schematic of the transcriptome sequencing workflow following METTL3 knockdown in MB49 cells. (B) Heatmap of differentially expressed genes after METTL3 knockdown (criteria for differential genes: p<0.05, fold change >1.5 or <0.67. (C) Volcano plot of differentially expressed genes after METTL3 knockdown. (D) Network diagram of GO enrichment analysis of differentially expressed genes. (E) Chemokines with differential expression following METTL3 silencing. (F–G) Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis of CXCL5 and CCL5 mRNA expression levels following METTL3 overexpression or knockdown in MB49 cells. (H) RT-qPCR analysis of CXCL5 and CCL5 mRNA expression levels in MB49 cells treated with DMSO or STM2457 for 72 hours. (I) ELISA of CXCL5 and CCL5 secretion levels in the culture supernatant of MB49 cell lines; ELISA of CXCL5 and CCL5 levels in (J) mouse tumor tissues and (K) in peripheral blood serum. RT-qPCR analysis of CXCL5 and CCL5 mRNA expression levels in (L) 5637 cells and (M) T24 cells treated with DMSO or STM2457 for 72 hours. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: Secretion levels of CXCL5 and CCL5 were measured using the Mouse CXCL5 ELISA Kit (Elabscience, #E-EL-M0471) and the Mouse RANTES ELISA Kit (Elabscience, #E-EL-M0009), respectively.

Techniques: Expressing, Sequencing, Knockdown, Quantitative Proteomics, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Over Expression, Enzyme-linked Immunosorbent Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: METTL3 promotes an immunosuppressive microenvironment in bladder cancer via m6A-dependent CXCL5/CCL5 regulation

doi: 10.1136/jitc-2024-011108

Figure Lengend Snippet: METTL3 regulates CXCL5 expression through N6-methyladenosine (m6A) modification, thereby promoting myeloid-derived suppressor cell (MDSC) chemotaxis. (A) Schematic diagram of the workflow used for Methylated RNA Immunoprecipitation (MeRIP) sequencing and data analysis in METTL3-knockdown MB49 stable cell lines. (B) Bar chart showing the number of m6A modification sites identified in MeRIP sequencing results. (C) Venn diagram of downstream target gene screening for METTL3, intersecting chemokines significantly altered in RNA sequencing with those showing significant downregulation in m6A modification levels in m6A sequencing. (D) m6A peak map of CXCL5 mRNA modification sites. (E) Bar chart of MeRIP-qPCR results showing the m6A modification level of CXCL5 mRNA in MB49 cells after METTL3 knockdown. (F) RNA Immunoprecipitation (RIP) assay detecting the interaction between METTL3 and CXCL5 mRNA. (G) RNA degradation assay showing CXCL5 mRNA stability after silencing METTL3. (H) RNA degradation assay showing CXCL5 mRNA stability after treatment with METTL3 inhibitor STM2457 (2 µg/mL, 72 hours) in MB49 cells. (I) Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis of IGF2BP1 and CXCL5 mRNA expression levels in MB49 cells after silencing IGF2BP1. (J) RT-qPCR analysis of IGF2BP2 and CXCL5 mRNA expression levels in MB49 cells after silencing IGF2BP2. (K) RT-qPCR analysis of METTL3, IGF2BP1, and CXCL5 mRNA expression levels in MB49 cells after overexpression of METTL3 and/or silencing of IGF2BP1. (L) Schematic of the animal experiment. (M) Images of bladder cancer tumors in mice. (N) Growth curves of bladder cancer tumors in mice. (O) Tumor weights of bladder cancer tumors in mice. ns, no significance. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: Secretion levels of CXCL5 and CCL5 were measured using the Mouse CXCL5 ELISA Kit (Elabscience, #E-EL-M0471) and the Mouse RANTES ELISA Kit (Elabscience, #E-EL-M0009), respectively.

Techniques: Expressing, Modification, Derivative Assay, Chemotaxis Assay, Methylation, RNA Immunoprecipitation, Sequencing, Knockdown, Stable Transfection, RNA Sequencing, Degradation Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Over Expression

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: CXCL16 mediate the intercommunication between CRC cells and KCs. (A)RNA-seq gene expression heatmap in CT26 and CT26-LM group. (B)RNA-seq gene expression heatmap in CT26 and co-CT26 group. (C)Venn diagram of genes positively correlated with liver metastasis analyzed through transcriptome microarrays between parent CT26 cells and CT26-LM or CT26 and co-CT26 cells. (D)Heatmap of top 20 changed genes among parent CT26 cells, LM and co-cultured CT26 cells. (E)Heatmap of top 10 changed cytokine genes among parent CT26 cells, LM and co-cultured CT26 cells. (F)qRT-PCR analysis of the mRNA expression of CXCL14, IL24, CXCL16 and IL1A in CT26 and CT26-LM cells. (G)qRT-PCR analysis of the mRNA expression of CXCL14, IL24, CXCL16 and IL1A in CT26 and co-CT26 cells. (H)ELISA assay revealed the CXCL16 upregulated in culture supernatants from CT26-LM cells. (I)Western blot analysis showed the changes of CXCL16 protein level in CT26, CT26-LM and co-CT26 cells.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

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

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: CXCL16/PI3K/FOXO3a signaling pathway mediates M2 polarization of KCs. (A)Western blot showed the protein expression of M2 markers in different concentration of CXCL16. (B)Western blot showed the protein expression of M2 markers in different concentration of ML339. (C)Western blot analysis of M2 markers (CD206, ARG1) and PI3K/AKT1/FOXO3a pathway in ImKC when treated with CXCL16 (200ng/ml), LM-CM or ML339. (D)Western blot analysis of M2 markers (CD206, ARG1) and PI3K/AKT1/FOXO3a pathway in ImKC when treated with CXCL16 (200ng/ml), LM-CM or LY294002. (E)Identification of transcription factor FOXO3a binding site. (F)Prediction of FOXO3a binding sites in the CD206 promoter region using JASPAR. (G)The promoter sequence of CD206 contains 3 putative FOXO3a transcriptional factor binding sites. (H)Luciferase reporter assays for the activity of wild-type and mutant CD206 promoters. (I)Quantitative analysis of ChIP experiments performed on DNA samples precipitated with antibodies against FOXO3a and IgG using primers detecting FOXO3a binding sites on CD206 promoter.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: Western Blot, Expressing, Concentration Assay, Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: Intervention of CXCL16/CXCR6 for suppression of CT26-LM metastasis in vivo. (A)Schematic representation of BALB/c mice were treated with ML339 once every two days after spleen injection CT26. (B)Liver metastasis nodules between control(DMSO) group and ML339 group (metastatic tumor nodules shown by arrows). (C)Dotplot showed the number of metastatic tumors between control(DMSO) group and ML339 group. (D)Barplot showed the whole liver weight between control(DMSO) group and ML339 group. (E)F4/80 IHC staining and quantitative analysis of AOD in tumor; scale bar 50μm. (F)Relative mRNA expression of M2 markers and M1 markers in control(DMSO) and ML339 group liver tissue. (G-H)IF staining of ARG1 and CD206 in control(DMSO) and ML339 group liver tissue; scale bar 50μm.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: In Vivo, Injection, Control, Immunohistochemistry, Expressing, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: High metastatic tumor-derived CXCL16 mediates liver colonization metastasis by inducing Kupffer cell polarization via the PI3K/AKT/FOXO3a pathway

doi: 10.1016/j.neo.2025.101174

Figure Lengend Snippet: Diagrammatic sketch for CXCL16/PI3K/AKT1/FOXO3a activation in M2-KCs to promote MET of liver-metastatic cells.

Article Snippet: CXCL16 levels in cell culture media and mouse serum were quantified using a Mouse CXCL16 ELISA kit (Elabscience, China) according to the manufacturer's protocol.

Techniques: Activation Assay