Journal: Frontiers in Molecular Biosciences

Article Title: JAK-centric explainable few-shot gene-expression diagnosis framework for alopecia via MultiPLIER priors and relation-style set-to-set comparison

doi: 10.3389/fmolb.2025.1753206

Figure Lengend Snippet: RT-qPCR validation of the IL2RB/IL2RG–EOMES–GZMA cytotoxic/JAK axis in alopecia areata (AA). Relative mRNA levels of EOMES, GZMA, IL2RB, and IL2RG are significantly elevated in AA lesional scalp compared with healthy controls, whereas androgenetic alopecia (AGA) samples are comparable to controls, indicating that this cytotoxic/JAK module is transcriptionally active in AA but largely quiescent in AGA. Data are shown as mean ± SD. Statistical annotations: ** p < 0.01 vs. control; n.s., not significant (AGA vs. control).

Article Snippet: Equal amounts of protein were resolved by SDS–PAGE and transferred to PVDF membranes at 200 mA for 2 h. Membranes were blocked in 5% non-fat milk at room temperature for 2 h and incubated overnight at 4 ° C with primary antibodies against β -actin (1:4000; 20536-1-AP, Proteintech, China), EOMES (1:5000; 83945-5-RR, Proteintech, China), GZMA (1:500; 11288-1-AP, Proteintech, China), IL2RB (1:5000; 13602-1-AP, Proteintech, China), and IL2RG (1:500; 11409-1-AP, Proteintech, China).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Control

Journal: Frontiers in Molecular Biosciences

Article Title: JAK-centric explainable few-shot gene-expression diagnosis framework for alopecia via MultiPLIER priors and relation-style set-to-set comparison

doi: 10.3389/fmolb.2025.1753206

Figure Lengend Snippet: GSEA plots for key genes. (a) EOMES. (b) GZMA. (c) IL2RB. (d) IL2RG.

Article Snippet: Equal amounts of protein were resolved by SDS–PAGE and transferred to PVDF membranes at 200 mA for 2 h. Membranes were blocked in 5% non-fat milk at room temperature for 2 h and incubated overnight at 4 ° C with primary antibodies against β -actin (1:4000; 20536-1-AP, Proteintech, China), EOMES (1:5000; 83945-5-RR, Proteintech, China), GZMA (1:500; 11288-1-AP, Proteintech, China), IL2RB (1:5000; 13602-1-AP, Proteintech, China), and IL2RG (1:500; 11409-1-AP, Proteintech, China).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: JAK-centric explainable few-shot gene-expression diagnosis framework for alopecia via MultiPLIER priors and relation-style set-to-set comparison

doi: 10.3389/fmolb.2025.1753206

Figure Lengend Snippet: Western blot validation of the IL2RB/IL2RG–EOMES–GZMA cytotoxic/JAK axis at the protein level. Protein expression of EOMES, GZMA, IL2RB, and IL2RG is markedly upregulated in AA lesional scalp compared with healthy controls, concordant with the RT-qPCR results, whereas AGA-affected scalp shows no significant difference relative to controls. Representative immunoblots and densitometric quantification (normalized to β -actin) are shown. Data are presented as mean ± SD. Statistical annotations: ** p < 0.01 vs. control; n.s., not significant (AGA vs. control).

Article Snippet: Equal amounts of protein were resolved by SDS–PAGE and transferred to PVDF membranes at 200 mA for 2 h. Membranes were blocked in 5% non-fat milk at room temperature for 2 h and incubated overnight at 4 ° C with primary antibodies against β -actin (1:4000; 20536-1-AP, Proteintech, China), EOMES (1:5000; 83945-5-RR, Proteintech, China), GZMA (1:500; 11288-1-AP, Proteintech, China), IL2RB (1:5000; 13602-1-AP, Proteintech, China), and IL2RG (1:500; 11409-1-AP, Proteintech, China).

Techniques: Western Blot, Biomarker Discovery, Expressing, Quantitative RT-PCR, Control

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Genotypes shown include WT (black) and Il2rb Mut (red). (A)–(E) each represent three individual experiments. (A) Total body or spleen weight of 6- to 8-week littermates, n = 15 per genotype. (B) Serum cytokine levels of IL-2 and IL-15c (IL-15/15Rα) comparing mice at age 4 weeks (left, WT, n = 5; Il2rb Mut , n = 6) and 7 weeks (right, IL-2, n = 19 per genotype; IL-15c, n = 17 for WT and n = 18 for Il2rb Mut ). (C) Representative dot plot depicting Tregs defined as CD4 + CD25 + Foxp3 + for genotypes indicated (left). Percentage of Tregs from CD4 + T cells, n = 11 per genotype (right). (D) Representative dot plot depicting subpopulations of CD4 + or CD8 + T cells: naive, lower right quad; central memory (TCM), upper right quad; and effector memory (TEM), upper left quad, for genotypes indicated (left). Stacked graphs showing T cell distribution, n = 11 per genotype (right). (E) Absolute numbers of Tregs–CD4 + CD25 + Foxp3 + (left), n = 11 per genotype. Absolute numbers of TEM–CD44 + CD62L − cells (right), n = 7 per genotype. Data are shown as mean ± SEM. All p values were calculated by non-parametric Mann-Whitney test for a single factor; displayed as **** p ≤ 0.0001, *** p ≤ 0.001, ** p ≤ 0.01, and * p ≤ 0.05 or ns, non-significant.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: MANN-WHITNEY

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Genotypes shown include WT (black) and Il2rb Mut (red). (A)–(H) each represent 2 individual experiments. (A) Total CD122 expression depicted as median fluorescence intensity (MFI) in subpopulations of splenic CD4 + and CD8 + T from both genotypes; naive (circles) and TEM cells (diamonds), n = 12 for WT, n = 14 for Il2rb Mut ; isotype control antibody MFI is indicated by the gray dotted line. (B) CD122 cell-surface expression (purple) measured by ImageStream. Membrane mask was applied to the analysis of cells stained with CD122 clone 5H4 (cell surface + intracellular). Images compare CD4 + (green) to CD8 + (yellow) splenocytes from WT (top) and Il2rb Mut (bottom). (C) ImageStream quantification, with membrane intensity/area. (D) CD25 expression depicted as in (A), n = 8 for WT and n = 9 for Il2rb Mut . (E) Frequency of IL-2-, IFN-γ-, granzyme B-, and perforin-producing CD4 + and CD8 + splenic T cells, n = 7 per genotype. (F and G) Activation and exhaustion markers PD-1 (F) and KLRG1 (G) measured in CD4 + and CD8 + splenocytes from both genotypes; naive (circles) and TEM cells (diamonds), n = 12 for WT and n = 14 for Il2rb Mut . (H) Representative dot plot distinguishing BM subsets, specifically LSK (Sca-1 + c-Kit + ) cells (top right quad) and myeloid progenitors (MPs; top left quad). Graphs show percentage of LSK cells (left) and MP cells (right), n = 6 per genotype. Data are shown as mean ± SEM. All p values were calculated by non-parametric Mann-Whitney test for a single factor. ImageStream statistics calculated by unpaired parametric Welch’s correction are displayed as **** p ≤ 0.0001, *** p ≤ 0.001, ** p ≤ 0.01, and * p ≤ 0.05 or ns.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: Expressing, Fluorescence, Control, Membrane, Staining, Activation Assay, MANN-WHITNEY

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Mixed (WT:WT or Mut:WT at 97:3, into WT) bone marrow chimera (BMC) mouse cells and sera were harvested and analyzed 12–14 weeks post-transplant. Black open shapes indicate WT cells from Mut:WT BMCs (WT/Mut:WT). Red open shapes indicate Il2rb Mut cells from Mut:WT BMCs (Mut/Mut:WT). Data depict one representative experiment in each of (A)–(I). (A) Serum cytokine levels of IL-2, n = 3 for WT:WT and n = 4 for Mut:WT, and of IL-15c, n = 2 for WT:WT and n = 4 for Mut:WT, measured as in . (B) Total CD122 expression depicted as median fluorescence intensity (MFI) in subpopulations of splenic CD4 + and CD8 + T from both genotypes; naive (circles) and TEM cells (diamonds), n = 6 per genotype; isotype control antibody MFI is indicated by the gray dotted line. (C) CD25 expression depicted as in (B), n = 6 per genotype. (D) CD122 cell-surface expression (purple) measured by ImageStream. Membrane mask was applied to the analysis of cells stained with CD122 clone 5H4 (cell surface + intracellular). Images compare CD4 + (green) to CD8 + (yellow) splenocytes from WT (CD45.1) and Il2rb Mut (CD45.2) chimeric mice. (E) ImageStream quantification, with membrane intensity/area, for WT (black) and Il2rb Mut (red). (F) Frequency of IL-2-, IFN-γ-, granzyme B-, and perforin-producing CD4 + and CD8 + splenic T cells, n = 6 per genotype. (G and H) Activation and exhaustion markers PD-1 (G) and KLRG1 (H) measured in CD4 + and CD8 + splenocytes from both genotypes; naive (opened circles) and TEM cells (opened diamonds), n = 6 per genotype. (I) T cell distribution, n = 7 per genotype, over three experiments. Comparison of TEM cell numbers between genotypes, n = 4 per genotype (right). Data show one representative experiment. Data are shown as mean ± SEM. All p values were calculated by non-parametric Mann-Whitney test for a single factor. ImageStream statistics calculated by unpaired parametric Welch’s correction are displayed as **** p ≤ 0.0001, *** p ≤ 0.001, ** p ≤ 0.01, and * p ≤ 0.05 or ns.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: Expressing, Fluorescence, Control, Membrane, Staining, Activation Assay, Comparison, MANN-WHITNEY

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Analyses of splenocytes and serum from mice age 13 weeks. Genotypes include WT, n = 6 (black), and Il2rb Mut , n = 2 (red). Transferred Tregs are defined as CD4 + CD25 + GFP + . (A) Serum cytokine levels of IL-2 and IL-15c (IL-15/15Rα) were measured by ELISA. (B) Body weight of neonatal WT or Il2rb Mut mice with (right) and without (left) WT Treg transfer. (C) Spleen weight. (D) Representative dot plots depicting T cell subsets, CD4 + (left) and CD8 + (right). Naive, lower right quad; central memory (TCM), upper right quad; and effector memory (TEM), upper left quad, for both genotypes. (E) Average T cell distribution for all populations and genotypes. (F) Total CD122 expression depicted as median fluorescence intensity (MFI) in subsets of naive (circles) and TEM (diamonds) cells; isotype control antibody MFI is indicated by the gray dotted line. (G and H) Activation and exhaustion markers PD-1 (G) and KLRG1 (H) measured in naive (circles) and TEM (diamonds) cells. Data are shown as mean ± SEM. All p values were calculated by non-parametric Mann-Whitney test for a single factor, displayed as **** p ≤ 0.0001, *** p ≤ 0.001, ** p ≤ 0.01, and * p ≤ 0.05 or ns.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Fluorescence, Control, Activation Assay, MANN-WHITNEY

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Splenocytes were stimulated with IL-7 and increasing doses of IL-2 (top) and IL-15 (bottom). Genotypes shown include WT (black) and Il2rb Mut (red). Transferred Tregs are marked by CD4 + CD25 + GFP + . Bar graphs (left) quantify pSTAT5 response as a percentage of CD8 + T naive and TEM cells, based on the threshold set on unstimulated condition and isotype control for pSTAT5 antibody MFI per representative animal per genotype. (A) Intact animals, n = 3 per genotype. (B) Mut:WT-mixed BMCs, n = 6, with congenic WT (black) and Il2rb Mut (red) donors. (C) Foxp3 − GFP + Tregs transferred into neonatal WT ( n = 6) and Il2rb Mut ( n = 2). The bivariate (right), representing each subpopulation from both genotypes across three experiments, shows unstimulated vs. maximum IL-2 and IL-15c concentrations. See for p value calculations. The p values are indicated by letters (a)–(h) for each comparison performed per cytokine.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: Control, Comparison

Journal: Cell reports

Article Title: A hypomorphic Il2rb mutant mouse model recapitulates and reveals mechanisms of human T cell immune dysregulation in IL-2Rβ deficiency

doi: 10.1016/j.celrep.2025.115902

Figure Lengend Snippet: Splenocytes were stimulated with IL-7 and increasing doses of IL-2 (top) and IL-15 (bottom). Genotypes shown include WT (black) and Il2rb Mut (red). Transferred Tregs are marked by CD4 + CD25 + GFP + . Bar graphs (left) quantify pSTAT5 response as a percentage of CD8 + T naive and TEM cells, based on the threshold set on unstimulated condition and isotype control for pSTAT5 antibody MFI per representative animal per genotype. (A) Intact animals, n = 3 per genotype. (B) Mut:WT-mixed BMCs, n = 6, with congenic WT (black) and Il2rb Mut (red) donors. (C) Foxp3 − GFP + Tregs transferred into neonatal WT ( n = 6) and Il2rb Mut ( n = 2). See for p value calculations. The p values are indicated by letters (a)–(h) for each comparison performed per cytokine.

Article Snippet: Il2rb Mut mice young cohort T Cell Analysis , Mendeley Data , https://doi.org/10.17632/tkwmmd27jh.1.

Techniques: Control, Comparison