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: bioRxiv

Article Title: PTPN22 interacts with EB1 to regulate T cell receptor signaling

doi: 10.1101/481507

Figure Lengend Snippet: A Compared with the PTPN22 WT, PTPN22 ΔP1 can increase remarkably the phosphorylation levels of ZAP-70, Lck and Erk. B, C, D Western blot and qPCR assays were used to test the protein and mRNA expression levels of CD25, CD69 and IL-2, and the results shown that the protein and mRNA expression levels of CD25, CD69 and IL-2 increased significantly in PTPN22 ΔP1 transfected cells. Western blot assays were used to test the protein expression levels of CD25 and CD69 (B-C) or qPCR assays were used to test the mRNA expression levels of CD25, CD69 and IL-2 (D). E, F, G Deletion of P1 domain of PTPN22 will not affect its enzyme activity. pNPP hydrolysis with PTPN22 catalytic domain and PTPN22 ΔP1 (E) or Kinetics parameters for the wild-type and the mutants of PTPN22 toward pNPP (F,G). Data information: All Experiments were carried three times (A-E) with similar results. In (A-C), Significant differences were calculated with t-test and are indicated with *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: CD25 ELISA was performed using the pre-packaged CD25 ELISA kit (DY2438, R & D system), while CD69 ELISA was performed using the pre-packaged CD69 ELISA kit (TWp022633, www.tw-reagent.com ) following the manufacturer’s protocols.

Techniques: Western Blot, Expressing, Transfection, Activity Assay

Journal: bioRxiv

Article Title: PTPN22 interacts with EB1 to regulate T cell receptor signaling

doi: 10.1101/481507

Figure Lengend Snippet: A, B Test the protein expression levels of CD25 and CD69 by use of ELISA. ELISA assays were used to test the expression levels of CD25 (A) or Same assays were used to test the expression levels of CD69 (B). C, D Test the mRNA transcription levels of CD25 and CD69 by use of qPCR. qPCR assays were used to test the expression levels of CD25 (C) or Same assays were used to test the expression levels of CD69 (D). Data information: Significant differences were calculated with t-test and are indicated with *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: CD25 ELISA was performed using the pre-packaged CD25 ELISA kit (DY2438, R & D system), while CD69 ELISA was performed using the pre-packaged CD69 ELISA kit (TWp022633, www.tw-reagent.com ) following the manufacturer’s protocols.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Multiple intravenous injections of allogeneic equine mesenchymal stem cells do not induce a systemic inflammatory response but do alter lymphocyte subsets in healthy horses

doi: 10.1186/s13287-015-0050-0

Figure Lengend Snippet: Multiple allogeneic mesenchymal stem cell (MSC) injections result in changes in splenic regulatory T cell percentages. (A-D) There were no significant changes in splenic CD21 + B-cell (A) , CD4 + T-cell (B) , or CD8 + T-cell percentages (C) or CD4/CD8 ratios (D) following multiple MSC injections. (E) There were no significant changes in activated (CD25 + ) lymphocyte proportions. (F) There were significantly higher percentages of splenic FoxP3 + regulatory T cells in the horses injected with bone marrow (BM)-derived MSCs compared with horses injected with adipose tissue (AT)-derived MSCs. Data are presented as mean ± standard error of the mean. * P <0.05.

Article Snippet: The following antibodies were used: mouse-anti-equine CD3 (clone UC F6G 1:250; Jeffery Stott, University of California, Davis, CA, USA) [ ], mouse-anti-human CD21 (clone B-ly4 1:20; BD Pharmingen, San Jose, CA, USA) [ , ], polyclonal goat-anti-human CD25 (clone AF-223; R&D Systems) [ ], rat-anti-mouse/human FoxP3 (clone PCH101; ebioscience, San Diego, CA, USA) [ ], mouse-anti-CD4 (clone HB61A 1:133; VMRD, Pullman, WA, USA) [ ], mouse-anti-equine CD8 (clone F18P 1:500; J. Stott) [ ], and a donkey-anti-mouse secondary when necessary (1:50; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA).

Techniques: Injection, Derivative Assay