anti cxcl5  (R&D Systems)

Journal: Cell Reports Medicine

Article Title: Robust transcriptomic hallmarks targeting intratumor heterogeneity in intrahepatic cholangiocarcinoma

doi: 10.1016/j.xcrm.2026.102708

Figure Lengend Snippet: Distinct immune microenvironment across LIHV-defined subgroups (A) Immune cell abundance comparison across subgroups using the indicated immune analysis tools across four independent iCCA cohorts. Circle color and size represent log 2 fold change and p value, respectively. (B) Boxplots of CD66b + neutrophils, CD68 + macrophages, CD3 + T cells, CD20 + B cells, and αSMA + fibroblasts across subgroups (Mann-Whitney U test). (C) Heatmap of immune signatures and checkpoint expression summarized as mean Z scores per subgroup across four cohorts. ∗FDR < 0.05; ∗∗FDR < 0.01; ∗∗∗FDR < 0.001. (D) Heatmap of Spearman correlations between chemokine expression and neutrophil infiltration. IHC, immunohistochemistry. (E) Boxplot comparing CXCL5 expression across subgroups in the Fu-iCCA cohort (Mann-Whitney U test). (F) Dot heatmap of chemokine gene expression across major cell types in Xue’s scRNA-seq dataset. (G) Boxplots of average CXCL5 expression in tumor cells and macrophages across subgroups (Mann-Whitney U test). (H) Western blot validating CXCL5 knockdown and overexpression efficiency in RBE and HuCCT1 cells. (I) Quantification of migrated neutrophils in transwell assays co-cultured with modified RBE and HuCCT1 cells ( n = 4 replicates per group; mean ± SD; Student’s t test). (J) Western blot analysis of CXCL5 overexpression in KTP cells. (K) Tumor growth curves of mice injected with control or CXCL5-overexpressing KTP cells ( n = 6 per group; mean ± SEM; Student’s t test). (L) Proportion and number of infiltrated neutrophils in control and CXCL5-overexpressing KTP tumors ( n = 6 per group; Student’s t test). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. See also and and .

Article Snippet: A CXCL5 neutralizing antibody (R&D Systems, AF254) was added at 5 μg/mL.

Techniques: Comparison, MANN-WHITNEY, Expressing, Immunohistochemistry, Gene Expression, Western Blot, Knockdown, Over Expression, Cell Culture, Modification, Injection, Control

Journal: Cell Reports Medicine

Article Title: Enhancing gastric cancer immunotherapy: Insights from multi-omics analysis and innovations in photodynamic-chemotherapy nanoplatforms

doi: 10.1016/j.xcrm.2026.102635

Figure Lengend Snippet: Multi-omics analyses have revealed YAP1-driven immunosuppressive cellular communities in GC (A) Workflow of spatial transcriptomic profiling of GC specimens ( n = 17). (B) Classification of tumor spots in spatial transcriptome slides based on H&E staining and gene expression profiles. (C) Spatial feature plots illustrating YAP1 expression and the distribution of cell clusters in sample 1. (D) Abundance of cell types in spots with high versus low YAP1 expression, with differences tested by two-tailed t tests. Green p values indicate enrichment in YAP1 Low spots, and red p values indicate enrichment in YAP1 High spots. Cohen’s d applied to standardized differences in means of cell types between YAP1 Low and YAP1 High spots. (E) Overview of YAP1 expression analysis and clinical correlation in FFPE samples from 253 GC patients (cohort 4). (F) Quantitative comparison of CD8 + , CD4 + , and FOXP3 + immune cell densities between YAP1 Low and YAP1 High tumors in cohort 4 (Mann-Whitney U test; Spearman correlation). (G) Representative multiplex immunofluorescence (mIF) images of YAP1 (green), CD68 (red), SPP1 (yellow), and panCK (white) with DAPI (blue) and quantification of SPP1 + macrophage density relative to YAP1 H-scores. Scale bars, 100 μm. (H) Correlation heatmap of YAP1 and chemokine gene expression in tumor cells from cohort 1 scRNA-seq data. (I) Visualization of spatial transcriptomic slides showing co-expression of YAP1 (yellow) with CXCL3/5/8 and CCL20 (blue), with spots of co-expression indicated in green. (J) Comparative analysis of CXCL3/5/8 and CCL20 expression between YAP1 Low and YAP1 High spots within the tumor regions (Mann-Whitney U test). (K) Representative mIF images showing co-localization of YAP1 (green), CXCL5 (red), and panCK (white) with DAPI (blue) in tumor regions, accompanied by single-cell correlation analysis between YAP1 and CXCL5 mean fluorescence intensity (MFI) values. Scale bars, 100 μm. (L) Experimental schematic illustrating the co-culture setup of YAP1-overexpressing or YAP1-knockout HGC-27 cells with THP-1-derived macrophages, with selected conditions including CXCL5 neutralization, CXCR2 inhibition, and recombinant CXCL5 treatment. (M) Western blot analysis of CXCL5 protein expression in control, YAP1 OE, and YAP1 KO gastric cancer cell lines. (N) ELISA quantification of CXCL5 secretion in conditioned media from control, YAP1 OE, and YAP1 KO cells. (O) FACS quantification of SPP1 + CD68 + macrophages following co-culture with tumor cells under the indicated conditions ( YAP1 OE/KO, CXCL5 neutralization, CXCR2 inhibition [SB225002], and recombinant CXCL5 supplementation) (see also C).

Article Snippet: CXCL5 Protein, Human , MCE , HY-P7158.

Techniques: Biomarker Discovery, Staining, Gene Expression, Expressing, Two Tailed Test, Comparison, MANN-WHITNEY, Multiplex Assay, Immunofluorescence, Single Cell, Fluorescence, Co-Culture Assay, Knock-Out, Derivative Assay, Neutralization, Inhibition, Recombinant, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: Enhancing gastric cancer immunotherapy: Insights from multi-omics analysis and innovations in photodynamic-chemotherapy nanoplatforms

doi: 10.1016/j.xcrm.2026.102635

Figure Lengend Snippet: Efficient internalization of M@O-VNPs triggered dual ICD effects in vitro (A) Confocal laser scanning microscopy (CLSM) images and FACS analysis showing time-dependent uptake of FITC-labeled O-VNPs and M@O-VNPs by HGC-27 cells at 0.5, 2, and 4 h ( n = 3, scale bars, 20 μm). F-actin is labeled with phalloidin (red). (B) Transmission electron microscopy (TEM) images of HGC-27 cells 1 h after the uptake of O-VNPs and M@O-VNPs (scale bars, 1 μm). (C) Immunofluorescence images demonstrating YAP1 inhibition in HGC-27 cells after 24-h treatment with PBS, HMSN, VP, photoactivated VP, M@O-VNPs, and photoactivated M@O-VNPs. YAP1 is labeled in green, and F-actin is labeled in gray (scale bars, 20 μm). (D) CCK8 assay results showed the inhibition rates of HMSN, O-VNPs, M@O-VNPs, and photoactivated M@O-VNPs in HGC-27 and HIEC-6 cells after 24 h of treatment ( n = 3). (E) FACS analysis of apoptosis/necrosis in HGC-27 cells after 24-h treatment with PBS, free OXA, free OXA plus VP, and M@O-VNPs for 24 h, with and without laser irradiation, using Annexin V-FITC and PI staining ( n = 3). (F) Detection of reactive oxygen species (ROS) production using the DCFH-DA probe in HGC-27 cells treated with PBS, HMSN, free VP, free VP plus OXA, and M@O-VNPs for 24 h, with and without laser irradiation ( n = 3, scale bars = 50 μm). (G) CLSM analysis of immunogenic cell death (ICD) markers calreticulin (CRT) and HMGB1 in HGC-27 cells treated with PBS, HMSN, free VP, free VP plus OXA, and M@O-VNPs for 24 h, with and without laser irradiation ( n = 3, scale bars, 20 μm). (H) Schematic illustration of the Transwell co-culture system, in which murine splenocytes were incubated with YTN-16 GC cells for 48 h under different treatment conditions. (I–M) FACS analysis of immune cell subsets in mouse-spleen-derived cells co-cultured with YTN16 murine GC cells for 48 h. YTN16 cells were pretreated with different formulations for 24 h before co-culture. Quantified populations included mature DCs, CD8 + effector T cells, CD4 + T helper cells, Treg cells, and SPP1 + macrophages ( n = 3, two-tailed t test). Gating strategy and representative plots are shown in . ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (N) Measurement of CXCL5 secretion levels in co-culture supernatants under indicated treatments ( n = 3, two-tailed t test). ∗∗∗∗ p < 0.0001. (O) Schematic model depicting how photoactivated M@O-VNPs inhibit YAP1 expression, reduce CXCL5-CXCR2 signaling, and modulate SPP1 expression within the tumor-immune microenvironment.

Article Snippet: CXCL5 Protein, Human , MCE , HY-P7158.

Techniques: In Vitro, Confocal Laser Scanning Microscopy, Labeling, Transmission Assay, Electron Microscopy, Immunofluorescence, Inhibition, CCK-8 Assay, Irradiation, Staining, Co-Culture Assay, Incubation, Derivative Assay, Cell Culture, Two Tailed Test, Expressing