Journal: Cell reports
Article Title: Regulatory network analysis of Dclk1 gene expression reveals a tuft cell-ILC2 axis that inhibits pancreatic tumor progression
doi: 10.1016/j.celrep.2025.115734
Figure Lengend Snippet: (A) UMAP showing VIPER-inferred differential protein activity of IL-4RA in Dclk1 + cells. (B and C) Flow cytometry analysis for IL-13 expression in immune cells (CD45) isolated from normal pancreas and pancreas from Mist1-Kras mice and related quantification. (D and E) Flow cytometry analysis for EpCAM and ZsGreen of organoids from pancreas of Kras-Dclk1-DTR-ZsGreen mice in the presence of vehicle or IL-13 and related quantification. (F) RT-qPCR for the expression of Dclk1 . (G) RT-qPCR for the expression of Il13 in different immune cell populations in pancreas from Mist1-Kras mice: CD45 + immune cells, CD4 + T cells, γδ T cells, and ILC2s (Lin — CD45 + CD127 + CD90 + KLRG1 + ). (H and I) Flow cytometry analysis for ILC2s in normal and Mist1-Kras mice and related quantification. (J and K) ZsGreen expression in organoids from Kras-Dclk1-DTR-ZsGreen pancreas cultured with or without ILC2s, with quantification. (L) RT-qPCR for the expression of Dclk1 in organoids. (M) IF for ZsGreen of pancreas from Mist1-Kras-Dclk1-DTR-ZsGreen mice treated with control IGG or anti-IL-13 blocking antibody for 8 weeks. (N and O) Flow cytometry analysis for EpCAM and ZsGreen and related quantification. (P–R) RT-qPCR for the expression of Dclk1 , Cd24a , and Spib . Scale bars: 100 μm. Means ± SD. Statistical significance was evaluated by Student’s t test; * p ≤ 0.05, ** p ≤ 0.01, and **** p ≤ 0.001; n.s., not significant.
Article Snippet: For experiments involving viral infections, single cell suspensions in Matrigel were added with Spib overexpressing lentivirus (VectorBuilder) or Adeno-CMVCre (University of Iowa) at 500 PFU/cell and sub-sequently grown as described above.
Techniques: Activity Assay, Flow Cytometry, Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Control, Blocking Assay