Review



dap5 mice  (Jackson Laboratory)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 86
      Buy from Supplier

    Structured Review

    Jackson Laboratory dap5 mice
    Elevated <t>DAP5</t> expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Dap5 Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dap5+mice/pmc12915102-74-27-41?v=Jackson+Laboratory
    Average 86 stars, based on 1 article reviews
    dap5 mice - by Bioz Stars, 2026-06
    86/100 stars

    Images

    1) Product Images from "Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity"

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    Journal: Advanced Science

    doi: 10.1002/advs.202520625

    Elevated DAP5 expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Figure Legend Snippet: Elevated DAP5 expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Techniques Used: Expressing, Western Blot, Phospho-proteomics, Labeling, Comparison, Two Tailed Test

    Ablation of Dap5 in Tregs did not affect tTreg development and pTreg differentiation in vivo. a) Naïve CD4 + T cells were purified from 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice and differentiated into iTreg in vitro. The absence of Dap5 protein expression in iTregs from HO‐Dap5 ΔFoxp3 mice was assessed by performing WB. b) Similar spleen sizes between 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice. c) Bar plots showing unchanged proportions of indicated T cell subpopulations in the PBMC from 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. d) Dot plots displaying expression levels of the canonical marker genes across different thymocyte subpopulations. e) Bar plots showing unaltered proportions of Helios − Treg and Helios + Treg in PBMCs, spleen and lymph nodes between 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. f) Schematic diagram depicting the workflow of bone morrow (BM) chimera experiment. Briefly, the BM cells were isolated from age‐matched and sex‐matched Dap5 flox (CD45.2), HO‐Dap5 ΔFoxp3 (CD45.2) and wild type (WT) C57BL/6 (CD45.1). The BM cells from HO‐Dap5 ΔFoxp3 or Dap5 flox mice were mixed with BM cells from WT (CD45.1) mice at 1:1 ratio and intravenously injected into lethally irradiated (9 Gy) WT C57BL/6 recipient mice for subsequent bone marrow reconstitution. g) Bar plots comparing the contributions of Helios + Tregs and Helios − Tregs derived from donors of Dap5 flox and HO‐Dap5 ΔFoxp3 mice in the BM‐reconstituted recipients. p‐ values were determined by two‐tailed student's T ‐test (c, e and g), * p <0.05.
    Figure Legend Snippet: Ablation of Dap5 in Tregs did not affect tTreg development and pTreg differentiation in vivo. a) Naïve CD4 + T cells were purified from 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice and differentiated into iTreg in vitro. The absence of Dap5 protein expression in iTregs from HO‐Dap5 ΔFoxp3 mice was assessed by performing WB. b) Similar spleen sizes between 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice. c) Bar plots showing unchanged proportions of indicated T cell subpopulations in the PBMC from 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. d) Dot plots displaying expression levels of the canonical marker genes across different thymocyte subpopulations. e) Bar plots showing unaltered proportions of Helios − Treg and Helios + Treg in PBMCs, spleen and lymph nodes between 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. f) Schematic diagram depicting the workflow of bone morrow (BM) chimera experiment. Briefly, the BM cells were isolated from age‐matched and sex‐matched Dap5 flox (CD45.2), HO‐Dap5 ΔFoxp3 (CD45.2) and wild type (WT) C57BL/6 (CD45.1). The BM cells from HO‐Dap5 ΔFoxp3 or Dap5 flox mice were mixed with BM cells from WT (CD45.1) mice at 1:1 ratio and intravenously injected into lethally irradiated (9 Gy) WT C57BL/6 recipient mice for subsequent bone marrow reconstitution. g) Bar plots comparing the contributions of Helios + Tregs and Helios − Tregs derived from donors of Dap5 flox and HO‐Dap5 ΔFoxp3 mice in the BM‐reconstituted recipients. p‐ values were determined by two‐tailed student's T ‐test (c, e and g), * p <0.05.

    Techniques Used: In Vivo, Purification, In Vitro, Expressing, Marker, Isolation, Injection, Irradiation, Derivative Assay, Two Tailed Test

    HO‐ Dap5 ΔFoxp3 mice displayed lethal autoimmune tolerance defects. a) Under the same housing condition, the lifespan of HO‐ Dap5 ΔFoxp3 mice were significantly shortened compared with Dap5 flox littermates. b) Around 6‐8 weeks‐old, HO ‐Dap5 ΔFoxp3 mice had enlarged lymphoid organs, scaly patches on the skin tissues of the head and ears, hair loss, and swollen toes. c) Left panel : 2D uniform manifold approximate projection (2D‐UMAP) plots comparing the distribution of T cell subpopulations in PBMCs between 6‐weeks‐old Dap5 flox and HO‐ Dap5 ΔFoxp3 mice; right panel : stacked bar plots comparing the compositions of naïve T cells and effector T cells between Dap5 flox and HO‐ Dap5 ΔFoxp3 mice. d) Enhanced TCR clonal expansion (left) and reduced Shannon diversity indexes of TCRs (right) in the peripheral T cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. e) 2D‐UMAP plots showing elevated presence of Il4 + basophils in the peripheral blood of HO‐ Dap5 ΔFoxp3 mice. f) Bar plots comparing the frequencies of indicated immune cell populations in the peripheral blood from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice and age‐matched Dap5 flox mice. g) 2D‐UMAP plots displaying reduced B cell presence in the peripheral blood of HO‐Dap5 ΔFoxp3 mice. h) Reduced BCR clonal expansion (left) and increased Shannon diversity indexes of BCR (right) in the peripheral B cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. i) The bar plot demonstrates a reduction of peripheral B cells in HO‐Dap5 ΔFoxp3 mice relative to the age‐matched Dap5 flox mice. j) The multi‐color immunofluorescence (mIF) pictures showing impaired GC architectures and reduced B cells numbers in the spleens from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice compared with that from age‐matched Dap5 flox mice. DAPI (blue), CD3 (green), CD19 (red). k) bar plot indicating decreased frequencies of peripheral Treg cells in HO‐Dap5 ΔFoxp3 mice compared to Dap5 flox mice. l) Bar plots showing pTregs, but not CD4 + T or CD8 + T cells, derived from derived from HO‐Dap5 ΔFoxp3 mice were apoptotic (PI + annexin V + ). m) Naïve CD4 + T cells from Dap5 flox or HO‐Dap5 ΔFoxp3 mice were intravenously injected into Rag1 −/− mice. Approximately 20 days later, enlarged spleens and lymph nodes were observed in HO‐Dap5 ΔFoxp3 mice. n) The survival curve showing that the lifespan of Rag1 −/− mice receiving intravenous injection of naïve CD4 + T cells from HO‐Dap5 ΔFoxp3 mice was significantly shortened. p‐ values were determined by Log‐rank test (a and n), or two‐tailed student's T ‐test (f, g, h, i, k and l), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Figure Legend Snippet: HO‐ Dap5 ΔFoxp3 mice displayed lethal autoimmune tolerance defects. a) Under the same housing condition, the lifespan of HO‐ Dap5 ΔFoxp3 mice were significantly shortened compared with Dap5 flox littermates. b) Around 6‐8 weeks‐old, HO ‐Dap5 ΔFoxp3 mice had enlarged lymphoid organs, scaly patches on the skin tissues of the head and ears, hair loss, and swollen toes. c) Left panel : 2D uniform manifold approximate projection (2D‐UMAP) plots comparing the distribution of T cell subpopulations in PBMCs between 6‐weeks‐old Dap5 flox and HO‐ Dap5 ΔFoxp3 mice; right panel : stacked bar plots comparing the compositions of naïve T cells and effector T cells between Dap5 flox and HO‐ Dap5 ΔFoxp3 mice. d) Enhanced TCR clonal expansion (left) and reduced Shannon diversity indexes of TCRs (right) in the peripheral T cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. e) 2D‐UMAP plots showing elevated presence of Il4 + basophils in the peripheral blood of HO‐ Dap5 ΔFoxp3 mice. f) Bar plots comparing the frequencies of indicated immune cell populations in the peripheral blood from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice and age‐matched Dap5 flox mice. g) 2D‐UMAP plots displaying reduced B cell presence in the peripheral blood of HO‐Dap5 ΔFoxp3 mice. h) Reduced BCR clonal expansion (left) and increased Shannon diversity indexes of BCR (right) in the peripheral B cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. i) The bar plot demonstrates a reduction of peripheral B cells in HO‐Dap5 ΔFoxp3 mice relative to the age‐matched Dap5 flox mice. j) The multi‐color immunofluorescence (mIF) pictures showing impaired GC architectures and reduced B cells numbers in the spleens from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice compared with that from age‐matched Dap5 flox mice. DAPI (blue), CD3 (green), CD19 (red). k) bar plot indicating decreased frequencies of peripheral Treg cells in HO‐Dap5 ΔFoxp3 mice compared to Dap5 flox mice. l) Bar plots showing pTregs, but not CD4 + T or CD8 + T cells, derived from derived from HO‐Dap5 ΔFoxp3 mice were apoptotic (PI + annexin V + ). m) Naïve CD4 + T cells from Dap5 flox or HO‐Dap5 ΔFoxp3 mice were intravenously injected into Rag1 −/− mice. Approximately 20 days later, enlarged spleens and lymph nodes were observed in HO‐Dap5 ΔFoxp3 mice. n) The survival curve showing that the lifespan of Rag1 −/− mice receiving intravenous injection of naïve CD4 + T cells from HO‐Dap5 ΔFoxp3 mice was significantly shortened. p‐ values were determined by Log‐rank test (a and n), or two‐tailed student's T ‐test (f, g, h, i, k and l), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Techniques Used: Immunofluorescence, Derivative Assay, Injection, Two Tailed Test

    Mice with heterozygous Dap5 deletion in Tregs displayed unaltered peripheral immune homeostasis but enhanced Teff response against subcutaneous tumors. a) WB results showing intermediate expression level of Dap5 in pTregs from HE‐ Dap5 ΔFoxp3 mice. b) Bar plot displaying unaltered splenic Treg frequencies in HE‐ Dap5 ΔFoxp3 mice. c) Curves of body weight changes for HE‐ Dap5 ΔFoxp3 and Dap5 flox mice under DSS‐induced colitis modeling. d) Curves of body weight changes for Rag1 −/− mice receiving adoptive transfer of naïve CD4 + T cells or naïve CD4 + T cells mixed with Tregs purified from HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. e) Suppressed MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice. f) Curves monitoring MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. g) Suppressed Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice. h) Curves monitoring Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. i) Violin plots comparing gene expressions between tumor‐infiltrating CD4 + Tconv from HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. j) Bar plots comparing infiltrations of IFN‐γ + CD4 + T cells in the MC38 tumors that grew in HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. k) RNA velocity streamlines projected onto the UMAP‐based embedding. Cells were grouped according to their annotations. l) The directed Partition‐based Graph Abstraction (PAGA) graph showing the connectivity of these CD8 + T subpopulations. The edge weights quantify the connectivity between cell groups. m) Projection of RNA velocity streamlines on UMAP, grouped by cell type. n) GSEA plots displaying enriched signaling pathways in tumor‐infiltrating CD8 + T cells from HE‐ Dap5 ΔFoxp3 mice in contrast to that from Dap5 flox mice. o) Bar plots comparing infiltrations of IFN‐γ + CD8 + T and GZMB + CD8 + T cells between HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. p‐ values were determined by two‐tailed student's T ‐test (b‐d, f, h, j and o), * p <0.05, ** p <0.01, **** p <0.001, *** p <0.001.
    Figure Legend Snippet: Mice with heterozygous Dap5 deletion in Tregs displayed unaltered peripheral immune homeostasis but enhanced Teff response against subcutaneous tumors. a) WB results showing intermediate expression level of Dap5 in pTregs from HE‐ Dap5 ΔFoxp3 mice. b) Bar plot displaying unaltered splenic Treg frequencies in HE‐ Dap5 ΔFoxp3 mice. c) Curves of body weight changes for HE‐ Dap5 ΔFoxp3 and Dap5 flox mice under DSS‐induced colitis modeling. d) Curves of body weight changes for Rag1 −/− mice receiving adoptive transfer of naïve CD4 + T cells or naïve CD4 + T cells mixed with Tregs purified from HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. e) Suppressed MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice. f) Curves monitoring MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. g) Suppressed Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice. h) Curves monitoring Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. i) Violin plots comparing gene expressions between tumor‐infiltrating CD4 + Tconv from HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. j) Bar plots comparing infiltrations of IFN‐γ + CD4 + T cells in the MC38 tumors that grew in HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. k) RNA velocity streamlines projected onto the UMAP‐based embedding. Cells were grouped according to their annotations. l) The directed Partition‐based Graph Abstraction (PAGA) graph showing the connectivity of these CD8 + T subpopulations. The edge weights quantify the connectivity between cell groups. m) Projection of RNA velocity streamlines on UMAP, grouped by cell type. n) GSEA plots displaying enriched signaling pathways in tumor‐infiltrating CD8 + T cells from HE‐ Dap5 ΔFoxp3 mice in contrast to that from Dap5 flox mice. o) Bar plots comparing infiltrations of IFN‐γ + CD8 + T and GZMB + CD8 + T cells between HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. p‐ values were determined by two‐tailed student's T ‐test (b‐d, f, h, j and o), * p <0.05, ** p <0.01, **** p <0.001, *** p <0.001.

    Techniques Used: Expressing, Adoptive Transfer Assay, Purification, Protein-Protein interactions, Two Tailed Test

    LLPS of DAP5. a) Phylogenetic tree plot showing that DAP5 is highly conserved in eukaryotic species. b) DAP5 domains aligned with the results of predictor of natural disordered regions (PONDR) and net charge per residue (NCPR) analyses. c) LLPS of purified recombinant eGFP‐DAP5 protein was observed in the buffer supplemented with serial concentrations of protein and NaCl under confocal microscopy. d) The fluorescence recovery after photobleaching (FRAP) curve illustrating the dynamic recovery of eGFP‐DAP5 + puncta in HeLa cells stimulated with 50 µM ETO for 12 h. e) HeLa cells were transfected with indicated DAP5 constructs, then exposed to 50 µM ETO for 12 h. Subcellular eGFP‐DAP5 distribution were subsequently visualized under fluorescence microscopy. f) Observation of eGFP‐Dap5 + puncta in iTregs generated from eGFP‐Dap5‐KI mice under confocal microscopy. g) Total RNAs purified from HeLa cells dose‐dependently enhanced the formation of eGFP‐DAP5 + puncta. p‐ values were determined by two‐tailed student's T ‐test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Figure Legend Snippet: LLPS of DAP5. a) Phylogenetic tree plot showing that DAP5 is highly conserved in eukaryotic species. b) DAP5 domains aligned with the results of predictor of natural disordered regions (PONDR) and net charge per residue (NCPR) analyses. c) LLPS of purified recombinant eGFP‐DAP5 protein was observed in the buffer supplemented with serial concentrations of protein and NaCl under confocal microscopy. d) The fluorescence recovery after photobleaching (FRAP) curve illustrating the dynamic recovery of eGFP‐DAP5 + puncta in HeLa cells stimulated with 50 µM ETO for 12 h. e) HeLa cells were transfected with indicated DAP5 constructs, then exposed to 50 µM ETO for 12 h. Subcellular eGFP‐DAP5 distribution were subsequently visualized under fluorescence microscopy. f) Observation of eGFP‐Dap5 + puncta in iTregs generated from eGFP‐Dap5‐KI mice under confocal microscopy. g) Total RNAs purified from HeLa cells dose‐dependently enhanced the formation of eGFP‐DAP5 + puncta. p‐ values were determined by two‐tailed student's T ‐test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Techniques Used: Residue, Purification, Recombinant, Confocal Microscopy, Fluorescence, Transfection, Construct, Microscopy, Generated, Two Tailed Test

    Transcripts bound by DAP5 undergo active translation in Tregs. a) Schematic diagram exhibiting the workflow for the combined RIP‐seq, Ribo‐seq and RNA‐seq experiments. b) Bubble plots displaying enriched GO and KEGG terms that were obtained by performing enrichment analysis with transcripts bound by DAP5 in iTregs. c) Integrative Genomics Viewer (IGV) tracks showing representative binding peaks of DAP5 that are associated with cell survival, proliferation and immunomodulation in human iTreg cells. d) Bar plots showing enriched presence of transcripts bound by DAP5 in iTregs. For histogram plotting, qRT‐PCR assays were performed with DAP5‐RIP assay elutes. The RIP‐qPCR data analysis was performed according to the ΔΔCt method. [ <xref ref-type= 77 , 78 ] Briefly, ΔCt [normalized RIP] = (Ct [RIP]‐(Ct [Input]‐Log 2 (Input Dilution Factor))), % Input = 2 (‐ΔCt[normalized RIP]) . e) Addition of Cy5‐labeled 5′UTR of MCL1 enhanced formation of eGFP‐DAP5 + puncta in the buffer containing 100 mM NaCl, 20 mM Tris‐HCl (pH = 7.5), 5% PEG8000 and 2 µM eGFP‐DAP5. Pictures were taken under confocal microscopy. f) The volcano plot depicting transcripts in iTregs with significantly increased translation efficiencies (TE) compared to naïve CD4 + T cells. TE values were determined by combined analysis of Ribo‐seq and RNA‐seq data. Transcripts meeting the criteria of absolute log 2 TE − iTreg TE − na ï ve CD 4 ≥ 2 and adjusted p‐ value < 0.05 were designated as differentially translated. g) Lower panel : the curves displaying the density distributions of transcripts along their relative TE values ( log 2 TE − iTreg TE − na ï ve CD 4 ) (X‐axis). Transcripts were grouped into three categories: DAP5‐bound and hypo‐m 6 A modified, DAP5‐bound and m 6 A modified, and m 6 A modified only; upper panel : box plots comparing TE values of the aforementioned categories of transcripts within three indicated ranges of relative TE values: log 2 TE − iTreg TE − na ï ve CD 4 <‐1, 1≤ log 2 TE − iTreg TE − na ï ve CD 4 <4.4, 4.4≤ log 2 TE − iTreg TE − na ï ve CD 4 ≤10. h) Bar plots displaying efficiencies of in vitro mouse iTreg differentiation induced by FB23‐2 or STM2457. i) Flow cytometric histograms comparing Ki‐67 expression levels between mouse iTregs induced by FB23‐2 and STM2457. P‐ values were determined by two‐tailed student's T ‐test (d and h) or Kolmogorov‐Smirnov test and Kruskal‐Wallis (g), * p <0.05, ** p <0.01, *** p <0.001. " title="Transcripts bound by DAP5 undergo active translation in Tregs. a) Schematic diagram ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Transcripts bound by DAP5 undergo active translation in Tregs. a) Schematic diagram exhibiting the workflow for the combined RIP‐seq, Ribo‐seq and RNA‐seq experiments. b) Bubble plots displaying enriched GO and KEGG terms that were obtained by performing enrichment analysis with transcripts bound by DAP5 in iTregs. c) Integrative Genomics Viewer (IGV) tracks showing representative binding peaks of DAP5 that are associated with cell survival, proliferation and immunomodulation in human iTreg cells. d) Bar plots showing enriched presence of transcripts bound by DAP5 in iTregs. For histogram plotting, qRT‐PCR assays were performed with DAP5‐RIP assay elutes. The RIP‐qPCR data analysis was performed according to the ΔΔCt method. [ 77 , 78 ] Briefly, ΔCt [normalized RIP] = (Ct [RIP]‐(Ct [Input]‐Log 2 (Input Dilution Factor))), % Input = 2 (‐ΔCt[normalized RIP]) . e) Addition of Cy5‐labeled 5′UTR of MCL1 enhanced formation of eGFP‐DAP5 + puncta in the buffer containing 100 mM NaCl, 20 mM Tris‐HCl (pH = 7.5), 5% PEG8000 and 2 µM eGFP‐DAP5. Pictures were taken under confocal microscopy. f) The volcano plot depicting transcripts in iTregs with significantly increased translation efficiencies (TE) compared to naïve CD4 + T cells. TE values were determined by combined analysis of Ribo‐seq and RNA‐seq data. Transcripts meeting the criteria of absolute log 2 TE − iTreg TE − na ï ve CD 4 ≥ 2 and adjusted p‐ value < 0.05 were designated as differentially translated. g) Lower panel : the curves displaying the density distributions of transcripts along their relative TE values ( log 2 TE − iTreg TE − na ï ve CD 4 ) (X‐axis). Transcripts were grouped into three categories: DAP5‐bound and hypo‐m 6 A modified, DAP5‐bound and m 6 A modified, and m 6 A modified only; upper panel : box plots comparing TE values of the aforementioned categories of transcripts within three indicated ranges of relative TE values: log 2 TE − iTreg TE − na ï ve CD 4 <‐1, 1≤ log 2 TE − iTreg TE − na ï ve CD 4 <4.4, 4.4≤ log 2 TE − iTreg TE − na ï ve CD 4 ≤10. h) Bar plots displaying efficiencies of in vitro mouse iTreg differentiation induced by FB23‐2 or STM2457. i) Flow cytometric histograms comparing Ki‐67 expression levels between mouse iTregs induced by FB23‐2 and STM2457. P‐ values were determined by two‐tailed student's T ‐test (d and h) or Kolmogorov‐Smirnov test and Kruskal‐Wallis (g), * p <0.05, ** p <0.01, *** p <0.001.

    Techniques Used: RNA Sequencing, Binding Assay, Quantitative RT-PCR, Labeling, Confocal Microscopy, Modification, In Vitro, Expressing, Two Tailed Test

    Dap5 promotes IL2RA and MCL1 translation to maintain ti‐Treg stability and survival. a) Volcano plots showing differentially expressed genes between ti‐Tregs from HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. b) 2D‐UMAP plots displaying reduced Foxp3 expression among ti‐Tregs from HE ‐Dap5 ΔFoxp3 mice. c) Bar plots comparing CD25 expressions in splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. d) Bar plots comparing frequencies of splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. e) Bar plots comparing proportions of splenic or tumor‐infiltrating cleaved‐caspase3 + Tregs between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. f) Immunoblotting results showing absence of Mcl‐1 expression in iTregs derived from HO‐ Dap5 ΔFoxp3 . g) Construction strategy for Dap5 flox Foxp3 CreERT2 strain. h) Curves monitoring MC38 tumor growth in Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. Tamoxifen was administered on the same day as tumor cell inoculation. i‐k) Bar plots comparing proportions of Tregs in CD4 + T cells (i), cleaved caspase‐3 + Tregs in total Tregs (j) and GZMB + CD8 + T in total CD8 + T cells (k) in tumors from Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. P‐ values were determined by two‐tailed student's T ‐test (c, f, h, i, j and k) * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Figure Legend Snippet: Dap5 promotes IL2RA and MCL1 translation to maintain ti‐Treg stability and survival. a) Volcano plots showing differentially expressed genes between ti‐Tregs from HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. b) 2D‐UMAP plots displaying reduced Foxp3 expression among ti‐Tregs from HE ‐Dap5 ΔFoxp3 mice. c) Bar plots comparing CD25 expressions in splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. d) Bar plots comparing frequencies of splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. e) Bar plots comparing proportions of splenic or tumor‐infiltrating cleaved‐caspase3 + Tregs between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. f) Immunoblotting results showing absence of Mcl‐1 expression in iTregs derived from HO‐ Dap5 ΔFoxp3 . g) Construction strategy for Dap5 flox Foxp3 CreERT2 strain. h) Curves monitoring MC38 tumor growth in Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. Tamoxifen was administered on the same day as tumor cell inoculation. i‐k) Bar plots comparing proportions of Tregs in CD4 + T cells (i), cleaved caspase‐3 + Tregs in total Tregs (j) and GZMB + CD8 + T in total CD8 + T cells (k) in tumors from Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. P‐ values were determined by two‐tailed student's T ‐test (c, f, h, i, j and k) * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Techniques Used: Expressing, Western Blot, Derivative Assay, Two Tailed Test

    Graphical abstract: Dap5 functions as a molecular switch of translation mode in ti‐Tregs. Chronic stresses in the TME trigger ISR, leading to eIF‐2a phosphorylation and a consequent impairment of CDT activity in Tregs. Meanwhile, ti‐Tregs engage Dap5 to sustain alternate mode of translation of CD25 and MCL‐1, which are critical for ti‐Treg lineage stability and survival in the harsh TME.
    Figure Legend Snippet: Graphical abstract: Dap5 functions as a molecular switch of translation mode in ti‐Tregs. Chronic stresses in the TME trigger ISR, leading to eIF‐2a phosphorylation and a consequent impairment of CDT activity in Tregs. Meanwhile, ti‐Tregs engage Dap5 to sustain alternate mode of translation of CD25 and MCL‐1, which are critical for ti‐Treg lineage stability and survival in the harsh TME.

    Techniques Used: Phospho-proteomics, Activity Assay



    Similar Products

    egfp dap5 ki mice  (Shanghai Model Organisms Center)
    86
    Shanghai Model Organisms Center egfp dap5 ki mice
    Egfp Dap5 Ki Mice, supplied by Shanghai Model Organisms Center, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dap5+mice/pm41457459-281-2-12?v=Shanghai+Model+Organisms+Center
    Average 86 stars, based on 1 article reviews
    egfp dap5 ki mice - by Bioz Stars, 2026-06
    86/100 stars
    		  Buy from Supplier
    dap5 mice  (Jackson Laboratory)
    86
    Jackson Laboratory dap5 mice
    Elevated <t>DAP5</t> expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.
    Dap5 Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dap5+mice/pmc12915102-74-27-41?v=Jackson+Laboratory
    Average 86 stars, based on 1 article reviews
    dap5 mice - by Bioz Stars, 2026-06
    86/100 stars
    		  Buy from Supplier

    Image Search Results


    Elevated DAP5 expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Elevated DAP5 expression in ti‐Tregs is associated with ti‐Treg infiltrations. a) Western blotting results comparing activities of AKT/mTOR signaling pathway across naïve CD4 + T, iTh1, iTh2, iTh17 and iTregs. b) Flow cytometric results confirming reduced phosphorylation levels of Akt, S6K and 4E‐BP1 in mouse splenic Tregs than in conventional T cells (Tconv). c) Flow cytometric results revealing increased mean fluorescent intensities (MFI) of p‐eIF‐2α and Atf4 in ti‐Tregs than in pTregs. d) Box plots displaying higher puromycin‐labeling intensities among ti‐Tregs than in pTregs. e) Left panel: the representative histograms depicting distribution of Dap5 expressions among pTregs and ti‐Tregs; right panel: dot plots showing pairwise comparison of Dap5 MFI between ti‐Tregs and pTregs. f) The MFI of DAP5 among tiTregs compared with that in Tregs from paracancerous tissues. The p ‐value was determined by performing two‐tailed paired T test. g) Scatter plot displaying positive correlation between the DAP5 MFI and ti‐Treg frequencies in CRCs. p‐ values were determined by two‐tailed paired Student's T ‐test (b, c, e and f), unpaired Student's T ‐test (d) and Spearman's correlation Test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Expressing, Western Blot, Phospho-proteomics, Labeling, Comparison, Two Tailed Test

    Ablation of Dap5 in Tregs did not affect tTreg development and pTreg differentiation in vivo. a) Naïve CD4 + T cells were purified from 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice and differentiated into iTreg in vitro. The absence of Dap5 protein expression in iTregs from HO‐Dap5 ΔFoxp3 mice was assessed by performing WB. b) Similar spleen sizes between 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice. c) Bar plots showing unchanged proportions of indicated T cell subpopulations in the PBMC from 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. d) Dot plots displaying expression levels of the canonical marker genes across different thymocyte subpopulations. e) Bar plots showing unaltered proportions of Helios − Treg and Helios + Treg in PBMCs, spleen and lymph nodes between 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. f) Schematic diagram depicting the workflow of bone morrow (BM) chimera experiment. Briefly, the BM cells were isolated from age‐matched and sex‐matched Dap5 flox (CD45.2), HO‐Dap5 ΔFoxp3 (CD45.2) and wild type (WT) C57BL/6 (CD45.1). The BM cells from HO‐Dap5 ΔFoxp3 or Dap5 flox mice were mixed with BM cells from WT (CD45.1) mice at 1:1 ratio and intravenously injected into lethally irradiated (9 Gy) WT C57BL/6 recipient mice for subsequent bone marrow reconstitution. g) Bar plots comparing the contributions of Helios + Tregs and Helios − Tregs derived from donors of Dap5 flox and HO‐Dap5 ΔFoxp3 mice in the BM‐reconstituted recipients. p‐ values were determined by two‐tailed student's T ‐test (c, e and g), * p <0.05.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Ablation of Dap5 in Tregs did not affect tTreg development and pTreg differentiation in vivo. a) Naïve CD4 + T cells were purified from 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice and differentiated into iTreg in vitro. The absence of Dap5 protein expression in iTregs from HO‐Dap5 ΔFoxp3 mice was assessed by performing WB. b) Similar spleen sizes between 4‐weeks‐old Dap5 flox and age‐matched HO‐Dap5 ΔFoxp3 mice. c) Bar plots showing unchanged proportions of indicated T cell subpopulations in the PBMC from 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. d) Dot plots displaying expression levels of the canonical marker genes across different thymocyte subpopulations. e) Bar plots showing unaltered proportions of Helios − Treg and Helios + Treg in PBMCs, spleen and lymph nodes between 4‐weeks‐old HO‐Dap5 ΔFoxp3 and age‐matched Dap5 flox mice. f) Schematic diagram depicting the workflow of bone morrow (BM) chimera experiment. Briefly, the BM cells were isolated from age‐matched and sex‐matched Dap5 flox (CD45.2), HO‐Dap5 ΔFoxp3 (CD45.2) and wild type (WT) C57BL/6 (CD45.1). The BM cells from HO‐Dap5 ΔFoxp3 or Dap5 flox mice were mixed with BM cells from WT (CD45.1) mice at 1:1 ratio and intravenously injected into lethally irradiated (9 Gy) WT C57BL/6 recipient mice for subsequent bone marrow reconstitution. g) Bar plots comparing the contributions of Helios + Tregs and Helios − Tregs derived from donors of Dap5 flox and HO‐Dap5 ΔFoxp3 mice in the BM‐reconstituted recipients. p‐ values were determined by two‐tailed student's T ‐test (c, e and g), * p <0.05.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: In Vivo, Purification, In Vitro, Expressing, Marker, Isolation, Injection, Irradiation, Derivative Assay, Two Tailed Test

    HO‐ Dap5 ΔFoxp3 mice displayed lethal autoimmune tolerance defects. a) Under the same housing condition, the lifespan of HO‐ Dap5 ΔFoxp3 mice were significantly shortened compared with Dap5 flox littermates. b) Around 6‐8 weeks‐old, HO ‐Dap5 ΔFoxp3 mice had enlarged lymphoid organs, scaly patches on the skin tissues of the head and ears, hair loss, and swollen toes. c) Left panel : 2D uniform manifold approximate projection (2D‐UMAP) plots comparing the distribution of T cell subpopulations in PBMCs between 6‐weeks‐old Dap5 flox and HO‐ Dap5 ΔFoxp3 mice; right panel : stacked bar plots comparing the compositions of naïve T cells and effector T cells between Dap5 flox and HO‐ Dap5 ΔFoxp3 mice. d) Enhanced TCR clonal expansion (left) and reduced Shannon diversity indexes of TCRs (right) in the peripheral T cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. e) 2D‐UMAP plots showing elevated presence of Il4 + basophils in the peripheral blood of HO‐ Dap5 ΔFoxp3 mice. f) Bar plots comparing the frequencies of indicated immune cell populations in the peripheral blood from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice and age‐matched Dap5 flox mice. g) 2D‐UMAP plots displaying reduced B cell presence in the peripheral blood of HO‐Dap5 ΔFoxp3 mice. h) Reduced BCR clonal expansion (left) and increased Shannon diversity indexes of BCR (right) in the peripheral B cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. i) The bar plot demonstrates a reduction of peripheral B cells in HO‐Dap5 ΔFoxp3 mice relative to the age‐matched Dap5 flox mice. j) The multi‐color immunofluorescence (mIF) pictures showing impaired GC architectures and reduced B cells numbers in the spleens from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice compared with that from age‐matched Dap5 flox mice. DAPI (blue), CD3 (green), CD19 (red). k) bar plot indicating decreased frequencies of peripheral Treg cells in HO‐Dap5 ΔFoxp3 mice compared to Dap5 flox mice. l) Bar plots showing pTregs, but not CD4 + T or CD8 + T cells, derived from derived from HO‐Dap5 ΔFoxp3 mice were apoptotic (PI + annexin V + ). m) Naïve CD4 + T cells from Dap5 flox or HO‐Dap5 ΔFoxp3 mice were intravenously injected into Rag1 −/− mice. Approximately 20 days later, enlarged spleens and lymph nodes were observed in HO‐Dap5 ΔFoxp3 mice. n) The survival curve showing that the lifespan of Rag1 −/− mice receiving intravenous injection of naïve CD4 + T cells from HO‐Dap5 ΔFoxp3 mice was significantly shortened. p‐ values were determined by Log‐rank test (a and n), or two‐tailed student's T ‐test (f, g, h, i, k and l), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: HO‐ Dap5 ΔFoxp3 mice displayed lethal autoimmune tolerance defects. a) Under the same housing condition, the lifespan of HO‐ Dap5 ΔFoxp3 mice were significantly shortened compared with Dap5 flox littermates. b) Around 6‐8 weeks‐old, HO ‐Dap5 ΔFoxp3 mice had enlarged lymphoid organs, scaly patches on the skin tissues of the head and ears, hair loss, and swollen toes. c) Left panel : 2D uniform manifold approximate projection (2D‐UMAP) plots comparing the distribution of T cell subpopulations in PBMCs between 6‐weeks‐old Dap5 flox and HO‐ Dap5 ΔFoxp3 mice; right panel : stacked bar plots comparing the compositions of naïve T cells and effector T cells between Dap5 flox and HO‐ Dap5 ΔFoxp3 mice. d) Enhanced TCR clonal expansion (left) and reduced Shannon diversity indexes of TCRs (right) in the peripheral T cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. e) 2D‐UMAP plots showing elevated presence of Il4 + basophils in the peripheral blood of HO‐ Dap5 ΔFoxp3 mice. f) Bar plots comparing the frequencies of indicated immune cell populations in the peripheral blood from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice and age‐matched Dap5 flox mice. g) 2D‐UMAP plots displaying reduced B cell presence in the peripheral blood of HO‐Dap5 ΔFoxp3 mice. h) Reduced BCR clonal expansion (left) and increased Shannon diversity indexes of BCR (right) in the peripheral B cells from HO‐ Dap5 ΔFoxp3 mice compared with that from Dap5 flox mice. i) The bar plot demonstrates a reduction of peripheral B cells in HO‐Dap5 ΔFoxp3 mice relative to the age‐matched Dap5 flox mice. j) The multi‐color immunofluorescence (mIF) pictures showing impaired GC architectures and reduced B cells numbers in the spleens from 6‐weeks‐old HO‐Dap5 ΔFoxp3 mice compared with that from age‐matched Dap5 flox mice. DAPI (blue), CD3 (green), CD19 (red). k) bar plot indicating decreased frequencies of peripheral Treg cells in HO‐Dap5 ΔFoxp3 mice compared to Dap5 flox mice. l) Bar plots showing pTregs, but not CD4 + T or CD8 + T cells, derived from derived from HO‐Dap5 ΔFoxp3 mice were apoptotic (PI + annexin V + ). m) Naïve CD4 + T cells from Dap5 flox or HO‐Dap5 ΔFoxp3 mice were intravenously injected into Rag1 −/− mice. Approximately 20 days later, enlarged spleens and lymph nodes were observed in HO‐Dap5 ΔFoxp3 mice. n) The survival curve showing that the lifespan of Rag1 −/− mice receiving intravenous injection of naïve CD4 + T cells from HO‐Dap5 ΔFoxp3 mice was significantly shortened. p‐ values were determined by Log‐rank test (a and n), or two‐tailed student's T ‐test (f, g, h, i, k and l), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Immunofluorescence, Derivative Assay, Injection, Two Tailed Test

    Mice with heterozygous Dap5 deletion in Tregs displayed unaltered peripheral immune homeostasis but enhanced Teff response against subcutaneous tumors. a) WB results showing intermediate expression level of Dap5 in pTregs from HE‐ Dap5 ΔFoxp3 mice. b) Bar plot displaying unaltered splenic Treg frequencies in HE‐ Dap5 ΔFoxp3 mice. c) Curves of body weight changes for HE‐ Dap5 ΔFoxp3 and Dap5 flox mice under DSS‐induced colitis modeling. d) Curves of body weight changes for Rag1 −/− mice receiving adoptive transfer of naïve CD4 + T cells or naïve CD4 + T cells mixed with Tregs purified from HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. e) Suppressed MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice. f) Curves monitoring MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. g) Suppressed Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice. h) Curves monitoring Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. i) Violin plots comparing gene expressions between tumor‐infiltrating CD4 + Tconv from HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. j) Bar plots comparing infiltrations of IFN‐γ + CD4 + T cells in the MC38 tumors that grew in HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. k) RNA velocity streamlines projected onto the UMAP‐based embedding. Cells were grouped according to their annotations. l) The directed Partition‐based Graph Abstraction (PAGA) graph showing the connectivity of these CD8 + T subpopulations. The edge weights quantify the connectivity between cell groups. m) Projection of RNA velocity streamlines on UMAP, grouped by cell type. n) GSEA plots displaying enriched signaling pathways in tumor‐infiltrating CD8 + T cells from HE‐ Dap5 ΔFoxp3 mice in contrast to that from Dap5 flox mice. o) Bar plots comparing infiltrations of IFN‐γ + CD8 + T and GZMB + CD8 + T cells between HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. p‐ values were determined by two‐tailed student's T ‐test (b‐d, f, h, j and o), * p <0.05, ** p <0.01, **** p <0.001, *** p <0.001.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Mice with heterozygous Dap5 deletion in Tregs displayed unaltered peripheral immune homeostasis but enhanced Teff response against subcutaneous tumors. a) WB results showing intermediate expression level of Dap5 in pTregs from HE‐ Dap5 ΔFoxp3 mice. b) Bar plot displaying unaltered splenic Treg frequencies in HE‐ Dap5 ΔFoxp3 mice. c) Curves of body weight changes for HE‐ Dap5 ΔFoxp3 and Dap5 flox mice under DSS‐induced colitis modeling. d) Curves of body weight changes for Rag1 −/− mice receiving adoptive transfer of naïve CD4 + T cells or naïve CD4 + T cells mixed with Tregs purified from HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. e) Suppressed MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice. f) Curves monitoring MC38 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. g) Suppressed Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice. h) Curves monitoring Panc02 tumor growth in HE‐ Dap5 ΔFoxp3 mice or Dap5 flox mice. i) Violin plots comparing gene expressions between tumor‐infiltrating CD4 + Tconv from HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. j) Bar plots comparing infiltrations of IFN‐γ + CD4 + T cells in the MC38 tumors that grew in HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. k) RNA velocity streamlines projected onto the UMAP‐based embedding. Cells were grouped according to their annotations. l) The directed Partition‐based Graph Abstraction (PAGA) graph showing the connectivity of these CD8 + T subpopulations. The edge weights quantify the connectivity between cell groups. m) Projection of RNA velocity streamlines on UMAP, grouped by cell type. n) GSEA plots displaying enriched signaling pathways in tumor‐infiltrating CD8 + T cells from HE‐ Dap5 ΔFoxp3 mice in contrast to that from Dap5 flox mice. o) Bar plots comparing infiltrations of IFN‐γ + CD8 + T and GZMB + CD8 + T cells between HE‐ Dap5 ΔFoxp3 and Dap5 flox mice. p‐ values were determined by two‐tailed student's T ‐test (b‐d, f, h, j and o), * p <0.05, ** p <0.01, **** p <0.001, *** p <0.001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Expressing, Adoptive Transfer Assay, Purification, Protein-Protein interactions, Two Tailed Test

    LLPS of DAP5. a) Phylogenetic tree plot showing that DAP5 is highly conserved in eukaryotic species. b) DAP5 domains aligned with the results of predictor of natural disordered regions (PONDR) and net charge per residue (NCPR) analyses. c) LLPS of purified recombinant eGFP‐DAP5 protein was observed in the buffer supplemented with serial concentrations of protein and NaCl under confocal microscopy. d) The fluorescence recovery after photobleaching (FRAP) curve illustrating the dynamic recovery of eGFP‐DAP5 + puncta in HeLa cells stimulated with 50 µM ETO for 12 h. e) HeLa cells were transfected with indicated DAP5 constructs, then exposed to 50 µM ETO for 12 h. Subcellular eGFP‐DAP5 distribution were subsequently visualized under fluorescence microscopy. f) Observation of eGFP‐Dap5 + puncta in iTregs generated from eGFP‐Dap5‐KI mice under confocal microscopy. g) Total RNAs purified from HeLa cells dose‐dependently enhanced the formation of eGFP‐DAP5 + puncta. p‐ values were determined by two‐tailed student's T ‐test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: LLPS of DAP5. a) Phylogenetic tree plot showing that DAP5 is highly conserved in eukaryotic species. b) DAP5 domains aligned with the results of predictor of natural disordered regions (PONDR) and net charge per residue (NCPR) analyses. c) LLPS of purified recombinant eGFP‐DAP5 protein was observed in the buffer supplemented with serial concentrations of protein and NaCl under confocal microscopy. d) The fluorescence recovery after photobleaching (FRAP) curve illustrating the dynamic recovery of eGFP‐DAP5 + puncta in HeLa cells stimulated with 50 µM ETO for 12 h. e) HeLa cells were transfected with indicated DAP5 constructs, then exposed to 50 µM ETO for 12 h. Subcellular eGFP‐DAP5 distribution were subsequently visualized under fluorescence microscopy. f) Observation of eGFP‐Dap5 + puncta in iTregs generated from eGFP‐Dap5‐KI mice under confocal microscopy. g) Total RNAs purified from HeLa cells dose‐dependently enhanced the formation of eGFP‐DAP5 + puncta. p‐ values were determined by two‐tailed student's T ‐test (g), * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Residue, Purification, Recombinant, Confocal Microscopy, Fluorescence, Transfection, Construct, Microscopy, Generated, Two Tailed Test

    Transcripts bound by DAP5 undergo active translation in Tregs. a) Schematic diagram exhibiting the workflow for the combined RIP‐seq, Ribo‐seq and RNA‐seq experiments. b) Bubble plots displaying enriched GO and KEGG terms that were obtained by performing enrichment analysis with transcripts bound by DAP5 in iTregs. c) Integrative Genomics Viewer (IGV) tracks showing representative binding peaks of DAP5 that are associated with cell survival, proliferation and immunomodulation in human iTreg cells. d) Bar plots showing enriched presence of transcripts bound by DAP5 in iTregs. For histogram plotting, qRT‐PCR assays were performed with DAP5‐RIP assay elutes. The RIP‐qPCR data analysis was performed according to the ΔΔCt method. [ <xref ref-type= 77 , 78 ] Briefly, ΔCt [normalized RIP] = (Ct [RIP]‐(Ct [Input]‐Log 2 (Input Dilution Factor))), % Input = 2 (‐ΔCt[normalized RIP]) . e) Addition of Cy5‐labeled 5′UTR of MCL1 enhanced formation of eGFP‐DAP5 + puncta in the buffer containing 100 mM NaCl, 20 mM Tris‐HCl (pH = 7.5), 5% PEG8000 and 2 µM eGFP‐DAP5. Pictures were taken under confocal microscopy. f) The volcano plot depicting transcripts in iTregs with significantly increased translation efficiencies (TE) compared to naïve CD4 + T cells. TE values were determined by combined analysis of Ribo‐seq and RNA‐seq data. Transcripts meeting the criteria of absolute log 2 TE − iTreg TE − na ï ve CD 4 ≥ 2 and adjusted p‐ value < 0.05 were designated as differentially translated. g) Lower panel : the curves displaying the density distributions of transcripts along their relative TE values ( log 2 TE − iTreg TE − na ï ve CD 4 ) (X‐axis). Transcripts were grouped into three categories: DAP5‐bound and hypo‐m 6 A modified, DAP5‐bound and m 6 A modified, and m 6 A modified only; upper panel : box plots comparing TE values of the aforementioned categories of transcripts within three indicated ranges of relative TE values: log 2 TE − iTreg TE − na ï ve CD 4 <‐1, 1≤ log 2 TE − iTreg TE − na ï ve CD 4 <4.4, 4.4≤ log 2 TE − iTreg TE − na ï ve CD 4 ≤10. h) Bar plots displaying efficiencies of in vitro mouse iTreg differentiation induced by FB23‐2 or STM2457. i) Flow cytometric histograms comparing Ki‐67 expression levels between mouse iTregs induced by FB23‐2 and STM2457. P‐ values were determined by two‐tailed student's T ‐test (d and h) or Kolmogorov‐Smirnov test and Kruskal‐Wallis (g), * p <0.05, ** p <0.01, *** p <0.001. " width="100%" height="100%">

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Transcripts bound by DAP5 undergo active translation in Tregs. a) Schematic diagram exhibiting the workflow for the combined RIP‐seq, Ribo‐seq and RNA‐seq experiments. b) Bubble plots displaying enriched GO and KEGG terms that were obtained by performing enrichment analysis with transcripts bound by DAP5 in iTregs. c) Integrative Genomics Viewer (IGV) tracks showing representative binding peaks of DAP5 that are associated with cell survival, proliferation and immunomodulation in human iTreg cells. d) Bar plots showing enriched presence of transcripts bound by DAP5 in iTregs. For histogram plotting, qRT‐PCR assays were performed with DAP5‐RIP assay elutes. The RIP‐qPCR data analysis was performed according to the ΔΔCt method. [ 77 , 78 ] Briefly, ΔCt [normalized RIP] = (Ct [RIP]‐(Ct [Input]‐Log 2 (Input Dilution Factor))), % Input = 2 (‐ΔCt[normalized RIP]) . e) Addition of Cy5‐labeled 5′UTR of MCL1 enhanced formation of eGFP‐DAP5 + puncta in the buffer containing 100 mM NaCl, 20 mM Tris‐HCl (pH = 7.5), 5% PEG8000 and 2 µM eGFP‐DAP5. Pictures were taken under confocal microscopy. f) The volcano plot depicting transcripts in iTregs with significantly increased translation efficiencies (TE) compared to naïve CD4 + T cells. TE values were determined by combined analysis of Ribo‐seq and RNA‐seq data. Transcripts meeting the criteria of absolute log 2 TE − iTreg TE − na ï ve CD 4 ≥ 2 and adjusted p‐ value < 0.05 were designated as differentially translated. g) Lower panel : the curves displaying the density distributions of transcripts along their relative TE values ( log 2 TE − iTreg TE − na ï ve CD 4 ) (X‐axis). Transcripts were grouped into three categories: DAP5‐bound and hypo‐m 6 A modified, DAP5‐bound and m 6 A modified, and m 6 A modified only; upper panel : box plots comparing TE values of the aforementioned categories of transcripts within three indicated ranges of relative TE values: log 2 TE − iTreg TE − na ï ve CD 4 <‐1, 1≤ log 2 TE − iTreg TE − na ï ve CD 4 <4.4, 4.4≤ log 2 TE − iTreg TE − na ï ve CD 4 ≤10. h) Bar plots displaying efficiencies of in vitro mouse iTreg differentiation induced by FB23‐2 or STM2457. i) Flow cytometric histograms comparing Ki‐67 expression levels between mouse iTregs induced by FB23‐2 and STM2457. P‐ values were determined by two‐tailed student's T ‐test (d and h) or Kolmogorov‐Smirnov test and Kruskal‐Wallis (g), * p <0.05, ** p <0.01, *** p <0.001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: RNA Sequencing, Binding Assay, Quantitative RT-PCR, Labeling, Confocal Microscopy, Modification, In Vitro, Expressing, Two Tailed Test

    Dap5 promotes IL2RA and MCL1 translation to maintain ti‐Treg stability and survival. a) Volcano plots showing differentially expressed genes between ti‐Tregs from HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. b) 2D‐UMAP plots displaying reduced Foxp3 expression among ti‐Tregs from HE ‐Dap5 ΔFoxp3 mice. c) Bar plots comparing CD25 expressions in splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. d) Bar plots comparing frequencies of splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. e) Bar plots comparing proportions of splenic or tumor‐infiltrating cleaved‐caspase3 + Tregs between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. f) Immunoblotting results showing absence of Mcl‐1 expression in iTregs derived from HO‐ Dap5 ΔFoxp3 . g) Construction strategy for Dap5 flox Foxp3 CreERT2 strain. h) Curves monitoring MC38 tumor growth in Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. Tamoxifen was administered on the same day as tumor cell inoculation. i‐k) Bar plots comparing proportions of Tregs in CD4 + T cells (i), cleaved caspase‐3 + Tregs in total Tregs (j) and GZMB + CD8 + T in total CD8 + T cells (k) in tumors from Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. P‐ values were determined by two‐tailed student's T ‐test (c, f, h, i, j and k) * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Dap5 promotes IL2RA and MCL1 translation to maintain ti‐Treg stability and survival. a) Volcano plots showing differentially expressed genes between ti‐Tregs from HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. b) 2D‐UMAP plots displaying reduced Foxp3 expression among ti‐Tregs from HE ‐Dap5 ΔFoxp3 mice. c) Bar plots comparing CD25 expressions in splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. d) Bar plots comparing frequencies of splenic or tumor‐infiltrating CD4 + Foxp3 + T cells between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. e) Bar plots comparing proportions of splenic or tumor‐infiltrating cleaved‐caspase3 + Tregs between HE‐ Dap5 ΔFoxp3 or Dap5 flox mice. f) Immunoblotting results showing absence of Mcl‐1 expression in iTregs derived from HO‐ Dap5 ΔFoxp3 . g) Construction strategy for Dap5 flox Foxp3 CreERT2 strain. h) Curves monitoring MC38 tumor growth in Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. Tamoxifen was administered on the same day as tumor cell inoculation. i‐k) Bar plots comparing proportions of Tregs in CD4 + T cells (i), cleaved caspase‐3 + Tregs in total Tregs (j) and GZMB + CD8 + T in total CD8 + T cells (k) in tumors from Dap5 fl/+ Foxp3 CreERT2 and Foxp3 CreERT2 mice. P‐ values were determined by two‐tailed student's T ‐test (c, f, h, i, j and k) * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Expressing, Western Blot, Derivative Assay, Two Tailed Test

    Graphical abstract: Dap5 functions as a molecular switch of translation mode in ti‐Tregs. Chronic stresses in the TME trigger ISR, leading to eIF‐2a phosphorylation and a consequent impairment of CDT activity in Tregs. Meanwhile, ti‐Tregs engage Dap5 to sustain alternate mode of translation of CD25 and MCL‐1, which are critical for ti‐Treg lineage stability and survival in the harsh TME.

    Journal: Advanced Science

    Article Title: Targeting DAP5 Disrupts Alternate Mode of Translational Initiation in Tregs and Potentiates Antitumor Immunity

    doi: 10.1002/advs.202520625

    Figure Lengend Snippet: Graphical abstract: Dap5 functions as a molecular switch of translation mode in ti‐Tregs. Chronic stresses in the TME trigger ISR, leading to eIF‐2a phosphorylation and a consequent impairment of CDT activity in Tregs. Meanwhile, ti‐Tregs engage Dap5 to sustain alternate mode of translation of CD25 and MCL‐1, which are critical for ti‐Treg lineage stability and survival in the harsh TME.

    Article Snippet: To dissect the function of Dap5 in Tregs in vivo, we generated transgenic mice with specific Dap5 ablation in Tregs ( Dap5 ΔFoxp3 ) by crossing the floxed‐ Dap5 mice with the Foxp3 ‐ Cre (B6.129(Cg)‐ Foxp3 4(YFP/icre)Ayr /J) strain from Jackson Laboratory that are widely used in Treg studies (Figure , Supporting Information).

    Techniques: Phospho-proteomics, Activity Assay