flt3 ligand  (R&D Systems)

Journal: eLife

Article Title: A single microRNA miR-195 rescues the arrested B cell development induced by EBF1 deficiency

doi: 10.7554/eLife.101510

Figure Lengend Snippet: ( A ) Flow cytometry analysis of control and Mir195 -expressing Lin – cells. HPCs from fetal livers of wild-type mice were cultured for 7 days on OP9 with SCF, Flt3-ligand, and IL-7, after infection with control or Mir195 retrovirus. Representative result of control (upper panel) and Mir195 (lower panel) viral infections is shown (n=3). ( B ) Outline of the in vitro culture system of Ebf1 -/- HPCs. ( C ) Flow cytometry analysis of control and Mir195 -expressing Ebf1 -/- HPCs. Shown data is representative of n=3. ( D ) Microarray analysis of Mir195 -expressing Ebf1 -/- HPCs. Log 2 fold-changes in the expression levels of genes related to B (left panel), T (middle-upper panel), NK (middle-lower panel), and myeloid (right panel) cell lineages were classified and are shown as colored columns. The analysis was carried out in duplicates.

Article Snippet: The infected and transduced Lin - cells were cultured and differentiated into B cells on OP9 cells in IMDM (Thermo Fisher) supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 0.1 mM non-essential amino acid solution, 50 μM 2-mercaptoethanol, 100 units/mL penicillin G, 100 μg/mL streptomycin (all from Wako), and 10 ng/mL recombinant SCF, IL-7, and Flt3-ligand (R&D Systems).

Techniques: Flow Cytometry, Control, Expressing, Cell Culture, Infection, In Vitro, Microarray

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