mouse pdgfra creer t2  (Jackson Laboratory)

Journal: iScience

Article Title: In vivo reprogramming of NG2 glia improves bladder function after spinal cord injury

doi: 10.1016/j.isci.2026.114850

Figure Lengend Snippet: SOX2-mediated glial reprogramming in the adult mouse spinal cord (A) Experimental scheme. SOX2-expressing lentiviruses were injected into the adult spinal cord, and tissues were analyzed 4 weeks (wk) later using immunohistochemistry (IHC). (B) Quantification of SOX2-induced DCX + immature neurons (mean ± SEM, n = 3–4 mice per group). Notably, DCX + cells were nearly absent in the p75-2-only condition. (C) Confocal images of DCX + cells induced by either wild-type SOX2 (SOX2 WT ) or the phospho-mimetic mutant SOX2 (SOX2 S251E ). Scale bars, 20 μm (D) Experimental scheme for tracing the cellular origin of mutant SOX2-reprogrammed cells. Adult transgenic mice were treated with tamoxifen (Tam), followed by intraspinal viral injections, and analyzed 4 weeks later. (E) Confocal images and quantifications (mean ± SEM, n = 3 mice per group) showing that NG2 glia are the cellular origin of mutant SOX2-reprogrammed cells in the adult spinal cord. NG2 glia and astrocytes were lineage-traced using Pdgfra-CreER T2 ;R26R-tdT and Aldh1l1-CreER T2 ;R26R-tdT mice, respectively. Scale bar, 20 μm (F) Experimental scheme for a time-course analysis of mutant SOX2-mediated reprogramming. dpv, days post virus injection. (G) Quantification of mutant SOX2-induced ASCL1 + neural progenitors (mean ± SEM, n = 3 mice per group). (H) Confocal images showing induction of ASCL1 + neural progenitors by mutant SOX2. Scale bar, 20 μm (I) Experimental scheme for tracing the progeny of mutant SOX2-induced ASCL1 + progenitors in the adult spinal cord. wpv, weeks post virus injection. (J) Quantification of mutant SOX2-induced tdT + cells at 4 wpv (mean ± SEM, n = 5 mice). (K) Confocal images showing robust detection of DCX + tdT + cells in mutant SOX2-expressing spinal cord. Scale bar, 20 μm.

Article Snippet: Mouse: Pdgfra-CreER T2 , The Jackson Laboratory , JAX: 032770; RRID: IMSR_JAX: 032770.

Techniques: Expressing, Injection, Immunohistochemistry, Mutagenesis, Transgenic Assay, Virus

Journal: Cancer Discovery

Article Title: Oncogenic KRAS-Dependent Stromal Interleukin-33 Directs the Pancreatic Microenvironment to Promote Tumor Growth

doi: 10.1158/2159-8290.CD-24-0100

Figure Lengend Snippet: Stromal IL33 promotes PDA growth. A, Genetic scheme of Pdgfra-CreER T2/+ ;Il33 f/f murine model. Tamoxifen induces activation of the Cre-ERT2 fusion protein, allowing recombination to occur. B, Experimental design for the Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumor model. OT, orthotopic, CreER = Pdgfra-CreER T2/+ , CreER;Il33 f/f = Pdgfra-CreER T2/+ ;Il33 f/f . C, Western blot of PDGFRα + cells sorted from CreER and CreER;Il33 f/f orthotopic tumors. Two tumors/mice were pooled in each lane. D, Relative and absolute tumor sizes from CreER and CreER;Il33 f/f orthotopic tumors. (E + F) Immunostainings of CreER and CreER;Il33 f/f tumors: E, = Co-IF staining of Ki67 (green), PDGFRα/β (red), E-Cadherin (white), and DAPI (blue), F, = IHC staining of Cleaved Caspase-3 (CC3). In quantification, each dot represents one animal. G, Treatment schedule for the Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumor model adapted for scRNA-seq. H, UMAP visualization of orthotopic scRNA-seq dataset split into CreER and CreER;Il33 f/f groups. I, Waterfall plot depicting differential pathway enrichment in tumor cells based on the Hallmark collection of annotations. Positive normalized enrichment scores are enriched in the control group. Pathways of interest are bolded. padj = Bonferroni-corrected P value. J, Violin plot depicting expression of Il1rl1 (ST2) in select leukocytes from scRNA-seq. K, Experimental design for Il1rl1 +/+ and Il1rl1 −/− orthotopic tumor experiment. L, Relative and absolute tumor sizes from Il1rl1 +/+ and Il1rl1 −/− orthotopic tumors. Tumor weight/body weight ratios are relative to the control group. Histogram data are mean ± standard deviation. Experiments with two conditions were compared using a two-tailed Student t test.

Article Snippet: To harvest PDGFRα + cells from Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumors for protein, single-cell suspensions were created as described above, and cells were labeled and subjected to CD140a (PDGFRα) magnetic selection as per the manufacturer’s instructions (#130-101-502, Miltenyi Biotec).

Techniques: Activation Assay, Western Blot, Staining, Immunohistochemistry, Control, Expressing, Standard Deviation, Two Tailed Test

Journal: Cancer Discovery

Article Title: Oncogenic KRAS-Dependent Stromal Interleukin-33 Directs the Pancreatic Microenvironment to Promote Tumor Growth

doi: 10.1158/2159-8290.CD-24-0100

Figure Lengend Snippet: Loss of stromal IL33 alters the ST2 + immune cell secretome, resulting in a shift in CAF differentiation. A, Gene expression of activation markers split by CreER and CreER;Il33 f/f from scRNA-seq. B, Waterfall plot depicting differential pathway enrichment in fibroblasts based on the Hallmark collection of annotations. Negative normalized enrichment scores are enriched in the experimental group. Pathways of interest are bolded. No genesets were enriched in the control group with Bonferroni-corrected P value (padj) of < 0.05. C, Chord diagram visualizing differentially enriched (Bonferroni-corrected P value < 0.05 and fold-change ≥0.25) predicted to interact with fibroblasts. Edge widths are proportional to predicted interaction strength. D, UMAP visualization of fibroblasts from the CreER and CreER;Il33 f/f scRNA-seq datasets. E, Gene expression of markers representing CAF subtypes. F, Il33 expression in each CAF population split by experimental group. G, Histogram depicting the frequency of each CAF population across the CreER and CreER;Il33 f/f scRNA-seq datasets. H, Chord diagram visualizing differentially enriched (Bonferroni-corrected P value < 0.05 and fold-change ≥0.25) fibroblast-derived ligands and their predicted interaction partners. Edge widths are proportional to predicted interaction strength.

Article Snippet: To harvest PDGFRα + cells from Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumors for protein, single-cell suspensions were created as described above, and cells were labeled and subjected to CD140a (PDGFRα) magnetic selection as per the manufacturer’s instructions (#130-101-502, Miltenyi Biotec).

Techniques: Gene Expression, Activation Assay, Control, Expressing, Derivative Assay

Journal: Cancer Discovery

Article Title: Oncogenic KRAS-Dependent Stromal Interleukin-33 Directs the Pancreatic Microenvironment to Promote Tumor Growth

doi: 10.1158/2159-8290.CD-24-0100

Figure Lengend Snippet: Inactivation of stromal IL33 enables cytotoxic T-cell activity. A, IHC staining of F4/80 in CreER and CreER ; Il33 f/f tumors. B, scRNA-seq gene expression of curated proinflammatory and immunosuppressive markers, grouped by cell type and split by experimental group. C, Chord diagram visualizing ligands differentially enriched (Bonferroni-corrected P value < 0.05 and fold-change ≥0.25) in CreER and CreER;Il33 f/f tumors that interact with CD8 + T cells. Edge widths are proportional to predicted interaction strength. Chemokines are bolded. D and E, Co-IF staining of CreER and CreER;Il33 f/f tumors: ( D ) = CD8 (green), Granzyme-B (red), E-Cadherin (white) and DAPI (blue), ( E ) = CD4 (yellow), Foxp3 (magenta), and DAPI (cyan). For staining quantification, each dot represents one animal, and values were compared using a two-tailed Student t test. Histogram data are mean ± standard deviation.

Article Snippet: To harvest PDGFRα + cells from Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumors for protein, single-cell suspensions were created as described above, and cells were labeled and subjected to CD140a (PDGFRα) magnetic selection as per the manufacturer’s instructions (#130-101-502, Miltenyi Biotec).

Techniques: Activity Assay, Immunohistochemistry, Gene Expression, Staining, Two Tailed Test, Standard Deviation

Journal: Cancer Discovery

Article Title: Oncogenic KRAS-Dependent Stromal Interleukin-33 Directs the Pancreatic Microenvironment to Promote Tumor Growth

doi: 10.1158/2159-8290.CD-24-0100

Figure Lengend Snippet: Expression of fibroblast IL33 is extrinsically induced by epithelial KRAS G12D and requires JAK1/2-STAT3 activation throughout tumorigenesis. A, Genetic scheme of the iKRAS G12D mouse. Doxycycline induces reversible expression of KRAS G12D in pancreatic epithelial cells. B, Diagram representing the various iKRAS G12D treatment models and collection points across tumorigenesis. Cae, caerulein; OT, orthotopic. C, UMAP visualization of iKRAS G12D scRNA-seq dataset. Projection on the left is colored by cell type (all datasets merged). Projections on the right are split by iKRAS G12D “ON” and “OFF” status and are colored by timepoint. D, Feature plot representation of Il33 expression levels split by iKRAS G12D “ON” and “OFF” status (all timepoints merged). E, Violin plots depicting fibroblast Il33 expression level per timepoint and split by iKRAS G12D “ON” and “OFF” status. Wilcoxon rank sum tests were performed between iKRAS G12D “ON” and “OFF” pairings per each timepoint, and Bonferroni adjusted P values are displayed above violins. F, GSEA enrichment plots of the Hallmark “IL6_JAK_STAT3_SIGNALING” pathway based on fibroblast iKRAS G12D “ON” and “OFF” differential gene expression analysis within each timepoint. G, Treatment scheme for iKRAS G12D “ON” model + JAK1/2 inhibitor. H, Co-IF staining of IL33 (green), PDGFRα/β (red), E-Cadherin (white), DAPI (blue). IL33 CTCF was quantified per individual ROI; each ROI encompasses one PDGFRα/β + cell. N = 3 mice were quantified per group. N in the figure represents the number of ROIs measured per group. P values represent a two-tailed Student t test. Line = Mean CTCF. I, Genetic scheme of Pdgfra-CreER T2/+ ; Stat3 f/f ( CreER;Stat3 f/f ) murine model. Tamoxifen induces activation of the Cre-ERT2 fusion protein, allowing recombination to occur. J, Diagram representing the treatment schedule for the CreER;Stat3 f/f orthotopic tumor model. K, Expression levels of Il33 in CAFs from J as measured by RT-qPCR. Values are normalized to Ppia (Cyclophilin A) and relative to the CreER group. Two-tailed Student t test was performed to compare groups; data are mean ± standard deviation.

Article Snippet: To harvest PDGFRα + cells from Pdgfra-CreER T2/+ ;Il33 f/f orthotopic tumors for protein, single-cell suspensions were created as described above, and cells were labeled and subjected to CD140a (PDGFRα) magnetic selection as per the manufacturer’s instructions (#130-101-502, Miltenyi Biotec).

Techniques: Expressing, Activation Assay, Gene Expression, Staining, Two Tailed Test, Quantitative RT-PCR, Standard Deviation