il34  (Miltenyi Biotec)

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: Analysis of IL34 expression in RCC patients (KIRC-TCGA and UroCCR cohorts) (A) Kaplan-Meier curves showing overall and progression-free survival in the KIRC-TCGA patient cohort. An optimal cutoff point was used to split the data into two groups on the basis of high and low IL34 gene expression. Log rank test, p values are indicated in the figure. (B and C) (B) Gene expression of IL34 in patient tumors stratified according to Fuhrman grade in the TCGA cohort. Median, min to max. Kruskal-Wallis test. (C) Gene expression of IL34 in patient tumors stratified according to the presence of distant metastases in the TCGA cohort. Median, min to max. Permutation Student’s t test. (D) (Left) Representative images of histological analysis of IL34 expression in RCC patient tumors from the UroCCR cohort. (Right) Quantification of the IL34 score stratified according to Fuhrman grade. Median, min to max. One-way ANOVA. Scale bar, 100 μm. (E) (Left) Representative images of IL34 expression in RCC patient tumors (UroCCR cohort). (Right) Quantification of the IL34 score on the basis of the absence or presence of distant metastases (M0 or M1, respectively). Median, min to max. Mann-Whitney U test. Scale bar, 100 μm. (F) Kaplan-Meier curves showing the progression-free survival of patients in the UroCCR cohort. The median value was used to stratify patients into two groups on the basis of high and low histological IL34 scores. Log rank test, p values are indicated on the figure. For all panels, number of patients per group and p values are indicated on the figure; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: Expressing, Gene Expression, MANN-WHITNEY

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: Single-nucleus RNA-seq of primary tumors and lung metastases generated from GFP-Renca cells (A) UMAP representation of the major cell types of 39,681 cells. (B) Heatmap showing the normalized expression of marker genes for each major cell type. (C) Fraction of cell type composition per library. PT, primary tumor; IVLM, intravenous-generated lung metastasis; LateLM, late-stage tumor-derived lung metastasis; EarlyLM, early-stage tumor-derived lung metastasis. (D) Abundancy of each cell type in control (Ctrl) and IL34-overexpressing (IL34-OE) Renca-injected mice. (E) Dot plot showing the expression of Il34, Csfr1, Trem2, Sdc1, and Ptprz1 per cell type. (F and G) (F) UMAP representation of the subclustering of the 7,207 cells in the immune compartment. (G) Heatmap showing the normalized expression of marker genes for each immune subtype. (H) Dot plot showing the expression of Csfr1, Adgre1, and Mrc1 per immune subtype. (I) Western blot analysis of CSF1R protein expression in murine Renca cancer cells and BMDMs.

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: RNA Sequencing, Generated, Expressing, Marker, Derivative Assay, Control, Injection, Western Blot

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: IL34 regulates the immune TME by accumulating monocyte-derived TAMs in primary tumors and lung metastases (A) RT-qPCR analysis of bulk RNA extracted from whole mouse renal tumors generated from IL34-OE (n = 5–7 mice per group) or Ctrl (n = 5–7 mice) Renca cells. Median, min to max. Mann-Whitney U test, ∗ p < 0.05, ∗∗∗ p < 0.001. (B) Immunohistochemistry (IHC) showing the expression of IL34, CSF1R, or F480 in Renca-generated orthotopic tumors. Scale bar, 100 μm. (C) IHC images of F4/80-positive cells in orthotopic xenografts generated from the human RCC cell lines 786-O and Caki2. Scale bar, 100 μm. (D) RT-qPCR analysis of bulk RNA extracted from metastatic lungs after tail vein injection of IL34-OE (n = 3–4 mice) or Ctrl ( n = 5 mice) Renca cells. Median, min to max. Mann-Whitney U test, ∗ p < 0.05. (E) IHC (left) and quantification (right) of F4/80-positive cells in lung metastases ( n = 3 mice per group). Scale bar, 100 μm; zoom, 50 μm. Mean ± SEM. Unpaired Student’s t tests, ∗ p < 0.05.

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: Derivative Assay, Quantitative RT-PCR, Generated, MANN-WHITNEY, Immunohistochemistry, Expressing, Injection

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: IL34-CSF1R axis is involved in MD-TAMs migration (A) Immunofluorescence analysis of proliferative MD-TAMs (i.e., Ki67 expression in CSF1R+ cells) in primary tumors generated from IL34-OE ( n = 5 mice) or Ctrl ( n = 6 mice) Renca cells. The graph on the right shows the quantification of total CSF1R+ cells. Scale bar, 100 μm; zoom, 50 μm. Mean ± SEM. Unpaired Student’s t tests, ∗ p < 0.05. (B) Quantification of proliferative MD-TAMs shown in (A). Mean ± SEM. Unpaired Student’s t tests. ns, non significant. FC, fold change. (C) Transwell migration assay of BMDM ( n = 6 mice per group) treated with either recombinant mouse IL34 or CSF1 (1 μg/mL, 24 hours). One dot represents a single-cell line of BMDM extracted from one mouse. Mean ± SEM. Kruskal-Wallis test, ∗∗∗ p < 0.001. (D) Transwell migration assay of BMDM after 24 h of coculture with Renca ( n = 7 mice), 786-O (n = 3–5 mice) or Caki2 (n = 4–6 mice) cells overexpressing mouse IL34 or an empty vector as a control under untreated or pexidartinib-treated (2.5 μM, 24 hours) conditions. One dot represents a single-cell line of BMDM extracted from one mouse. Mean ± SEM. two-way ANOVA, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (E) Histological analysis (left) and quantification (right) of protumor MD-TAMs by counting MRC1+ cells among F4/80+ cells ( n = 6 mice per group). Scale bar, 100 μm. Mean ± SEM. two-way ANOVA, ∗∗ p < 0.01.

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: Migration, Immunofluorescence, Expressing, Generated, Transwell Migration Assay, Recombinant, Plasmid Preparation, Control

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: Impact of the IL34-CSF1R axis in the immune-vascular crosstalk of RCC (A) Western blot analysis showing the levels of PD-L1 and VEGF expression in GFP-Renca cells and BMDM. (B) Histological analysis (left) and quantification (right) of immunosuppressive PD-L1+ F4/80+ MD-TAMs in lung metastases ( n = 6 mice per group). Scale bar, 100 μm. Mean ± SEM. two-way ANOVA, ∗∗∗ p < 0.001. (C) Primary tumor analysis revealed leaky vessels that were positive for endomucin and expressed low VE-cadherin, as indicated by white arrows ( n = 8 mice per group). Scale bar, 100 μm. Mean ± SEM. Mann-Whitney U test, ∗ p < 0.05. (D) Histological images of CSF1R+ cells associated with vessels (endomucin staining) in Renca-generated primary tumors (left) and quantification (right). Scale bar, 100 μm; zoom, 50 μm. Mean ± SEM ( n = 8 mice per group). Mann-Whitney U test, ∗∗∗ p < 0.001. (E) Quantification of extravasated Evans blue in Renca-generated primary tumors treated with pexidartinib (40 mg/kg) or placebo. Up to 12 mice per group from two independent experiments were analyzed. Data are expressed as the fold change compared with placebo control samples with tumor weight normalization. Mean ± SEM. two-way ANOVA, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (F) Histological images of leaky vessels (top) and quantification (bottom) of lung metastases from mice treated with pexidartinib (40 mg/kg) or placebo ( n = 6 per group). Scale bar, 50 μm. Mean ± SEM. two-way ANOVA, ∗ p < 0.05. (G) Volcano plot of the DEGs identified via MD-TAM. The x axis represents the log2-fold change in gene expression between the IL34-OE and Ctrl groups, indicating upregulation (right) and downregulation (left) in IL34-OE MD-TAM. The y axis represents the −log10 of the adjusted p value. The horizontal dashed line indicates the threshold for statistical significance (i.e., adjusted p value <0.05).

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: Western Blot, Expressing, MANN-WHITNEY, Staining, Generated, Control, Gene Expression

Journal: iScience

Article Title: Targeting the IL34-CSF1R axis improves metastatic renal cell carcinoma therapy outcome via immune-vascular crosstalk regulation

doi: 10.1016/j.isci.2025.112752

Figure Lengend Snippet: In RCC patients, high IL34 expression correlates with features of immunosuppression and a reduced response to immunotherapy (A) Gene Ontology term enrichment analysis of genes whose expression correlated with IL34 expression in KIRC-TCGA patients. (B) Spearman correlation analysis of IL34 expression with the MD-TAM markers CSF1R and CD68 or the exhausted T cell markers PDCD1 and CTLA4 in the KIRC-TCGA (left) and UroCCR (right) databases. (C) Kaplan-Meier survival analysis of patients stratified on the basis of IL34 expression (high vs. low) who received treatment (everolimus vs. nivolumab, CheckMate CM-025 cohort). Number of patients per group and p values are indicated in the figure.

Article Snippet: Il34 TaqMan Assay , ThermoFisher Scientific , Mm01243248_m1.

Techniques: Expressing