recombinant mouse scf c kit ligand  (R&D Systems)

Journal: Signal Transduction and Targeted Therapy

Article Title: Oncolytic adenovirus delivery of neoantigens sensitizes low-mutation tumors to anti-PD-1 therapy and prevents metastasis

doi: 10.1038/s41392-025-02511-5

Figure Lengend Snippet: Construction and in vivo validation of NeoViron. a ‒ d C57BL/6 mice inoculated with mICCN-4 or Hep53.4 cells were treated with PBS, vector or AdSVP-Flt3L. For the mICCN-4 model, treatment was initiated on day 8, with intratumoral injections of 1 × 10⁸ pfu OAV administered every other day for a total of five doses. For Hep53.4 tumors, treatment commenced on Day 6 with intratumoral injections of 2 × 10⁸ pfu OAV every other day for five doses. a , b Left, tumor images at the end point. Middle, tumor growth curves ( n = 6 mice per group). c , d CD103+ cDC1s were examined by flow cytometry ( c ) or immunofluorescence staining (MHC II+ CD103+ cells) ( d ). e Design of NeoViron, coexpressing neoantigens and Flt3L. f – h C57BL/6 mice inoculated with mICCN-4 were treated with PBS or 1 × 10⁸ pfu OAV (vector, AdSVP-NAg mICC , AdSVP-Flt3L or NeoViron) from Day 8 every other day for a total of five doses. f Left, experimental schematic. Middle, tumor images at the end point. Right, tumor growth curves ( n = 6 mice per group). g Percentages of PD1+ GZMB+ CD8+ T cells and PD1+ GZMB- CD8+ T cells infiltrating the tumor tissues of each group. h Representative immunofluorescence images of GZMB+ CD8+ T cells in tumor tissues. The data are shown as the means ± SDs ( a – g ) and are representative of two ( a – h ) independent experiments. Significance was calculated via one-way ANOVA ( a – c , g ) or two-way ANOVA ( a , b , f ). * P < 0.05, ** P < 0.01, *** P < 0.001. ns not significant

Article Snippet: One hundred milligrams of tumor tissue were finely sliced and incubated at 37 °C in 200 μL of PBS for 2 h. The supernatants were then collected and analyzed via a mouse Flt3L ELISA Kit (Proteintech, USA).

Techniques: In Vivo, Biomarker Discovery, Plasmid Preparation, Flow Cytometry, Immunofluorescence, Staining, Immunopeptidomics

Journal: Signal Transduction and Targeted Therapy

Article Title: Oncolytic adenovirus delivery of neoantigens sensitizes low-mutation tumors to anti-PD-1 therapy and prevents metastasis

doi: 10.1038/s41392-025-02511-5

Figure Lengend Snippet: NeoViron-induced CD8+ Trm cells prevent tumor metastasis. a ‒ g C57BL/6 mice inoculated with mICCN-4 or Hep53.4 cells were treated with PBS, vector or NeoViron/AdSVP-NAg Hep + AdSVP-Flt3L 5 times, followed by tumor excision on day 16. One week later, liver/lung metastasis models were established by injecting tumor cells via the spleen/tail vein. a , d Experimental schematic. b , e Tumor area of liver metastases and counts of lung metastases analyzed by ImageJ. c , f HE-stained images of liver and lung metastases at the end of the experiment. g Multiplex immunofluorescence images showing CD8+ Trm cells in liver and lung metastases. The data are shown as the means ± SDs ( b , e ) and are representative of two independent experiments ( a – h ). Significance was calculated via the Kruskal‒Wallis test ( b , e ). * P < 0.05, *** P < 0.001. ns not significant

Article Snippet: One hundred milligrams of tumor tissue were finely sliced and incubated at 37 °C in 200 μL of PBS for 2 h. The supernatants were then collected and analyzed via a mouse Flt3L ELISA Kit (Proteintech, USA).

Techniques: Plasmid Preparation, Staining, Multiplex Assay, Immunofluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Protein kinase Ds promote tumor angiogenesis through mast cell recruitment and expression of angiogenic factors in prostate cancer microenvironment

doi: 10.1186/s13046-019-1118-y

Figure Lengend Snippet: PKD2/3 enhance MCs migration through upregulation of SCF, CCL5 and CCL11 in prostate cancer cellsmRNA and the protein level of the scf , ccl5 and ccl11 were analyzed by real-time qPCR and ELISA in DU145 ( a ) and PC-3 M ( b ) cells transfected with siRNA of PKD2, PKD3. Data representing the means ± S.D. of three independent experiments was analyzed by one-way ANOVA for significance versus si-CTL. ***p < 0.001, **p < 0.01, *p < 0.05 versus si-CTL. c Knockdown efficiency of PKD2 and PKD3 in prostate cancer cells was verified by Western blotting. d DU145 cells were transfected with siRNA of PKD2, PKD3, the Conditional medium (CM) was collected to measure migration of P815 cells in response to SCF, CCL5, and CCL11 treatment by transwell assay. e Quantification were analyzed from data in D. ***p < 0.001 versus si-CTL by One-ANOVA tests

Article Snippet: Quantitative measurement of cytokines, including stem cell factor (SCF), Chemokine ligand 5 (CCL5), C-C motif chemokine 11(CCL11) and vascular endothelial growth factor (VEGF) secreted into conditioned medium was determined using ELISAs, according to the manufacturer’s protocol (BOSTER).

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Transfection, Knockdown, Western Blot, Transwell Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Protein kinase Ds promote tumor angiogenesis through mast cell recruitment and expression of angiogenic factors in prostate cancer microenvironment

doi: 10.1186/s13046-019-1118-y

Figure Lengend Snippet: PKD2 and PKD3 promote SCF, CCL5 and CCL11 expression through Erk1/2 signaling pathways. a-b Interaction of PKD2 or PKD3 with Erk1/2 was performed by co-IP assay in PC-3 M or DU145 prostate cancer cells. c-d DU145 cells ( c ) or PC-3 M cells ( d ) were transfected as indicated and treated with 100 nM PMA, phosphorylation and protein expression were detected by western blotting. e Overexpression efficiency of PKD2 and PKD3 in prostate cancer cells was verified by Western blotting. f ELISA were applied to measure SCF in conditional medium from DU145 cell transfected with GFP, GFP-PKD2, and GFP-PKD3 in present with or without Erk inhibitor PD98059(PD) treatment. g Real-time PCR was performed to analyze ccl5 expression in DU145 transfected with GFP, GFP-PKD2 and GFP-PKD3 plasmids followed by treatment with or without Erk inhibitor PD98059

Article Snippet: Quantitative measurement of cytokines, including stem cell factor (SCF), Chemokine ligand 5 (CCL5), C-C motif chemokine 11(CCL11) and vascular endothelial growth factor (VEGF) secreted into conditioned medium was determined using ELISAs, according to the manufacturer’s protocol (BOSTER).

Techniques: Expressing, Protein-Protein interactions, Co-Immunoprecipitation Assay, Transfection, Phospho-proteomics, Western Blot, Over Expression, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Protein kinase Ds promote tumor angiogenesis through mast cell recruitment and expression of angiogenic factors in prostate cancer microenvironment

doi: 10.1186/s13046-019-1118-y

Figure Lengend Snippet: PKD2 and PKD3 are required for SCF, CCL5, and CCL11 transcription. a Analysis of the binding site of p65(highlight in red) and AP1(highlight in blue) in the promoters of scf, ccl5 and ccl11 using UCSC software online. b Western blotting was used to ensure the knockdown effect. ChIP analysis of the binding of c-Jun ( c ), c-Fos (d) and NF-κB ( e ) to the scf, ccl5 and ccl11 gene promoter in PC-3 M cells depleted with siRNA of PKD2, PKD3. Student’s t-test , *p < 0.05, **p < 0.01 and ***p < 0.001 ( n = 3). Error bars indicate mean ± S.D.

Article Snippet: Quantitative measurement of cytokines, including stem cell factor (SCF), Chemokine ligand 5 (CCL5), C-C motif chemokine 11(CCL11) and vascular endothelial growth factor (VEGF) secreted into conditioned medium was determined using ELISAs, according to the manufacturer’s protocol (BOSTER).

Techniques: Binding Assay, Software, Western Blot, Knockdown

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Protein kinase Ds promote tumor angiogenesis through mast cell recruitment and expression of angiogenic factors in prostate cancer microenvironment

doi: 10.1186/s13046-019-1118-y

Figure Lengend Snippet: CRT0066101 reduces MCs recruitment and tumor angiogenesis in vivo. a Experimental setting. b C57BL6 mice bearing RM1 tumors were administered a daily vehicle [control group; 5% ( w / v ) dextrose] or CRT0066101 at 20 μM and 40 μM for 2 weeks (4 mice per group), then excised tumor images. c Tumor volume were represented at indicated day. d Immunohistochemistry staining for phospho-PKD, microvessel density (stained with CD31), and mast cells (stained with c-Kit). Representative image were shown in 400X under the microscope (Left panel). Quantification of the indicated parameter was analyzed among groups after treatment with CRT0066101 (Right panel). e Schematic model of the mechanistic role of PKD2 and PKD3 in tumor angiogenesis by regulating SCF-, CCL5-, and CCL11-mediated mast cell recruitment

Article Snippet: Quantitative measurement of cytokines, including stem cell factor (SCF), Chemokine ligand 5 (CCL5), C-C motif chemokine 11(CCL11) and vascular endothelial growth factor (VEGF) secreted into conditioned medium was determined using ELISAs, according to the manufacturer’s protocol (BOSTER).

Techniques: In Vivo, Control, Immunohistochemistry, Staining, Microscopy