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: Molecular Medicine Reports

Article Title: Expression, immunogenicity and clinical significance analysis of thyroid-stimulating hormone receptor fusion proteins

doi: 10.3892/mmr.2025.13639

Figure Lengend Snippet: Changes of specific regulatory T cell subsets in mice after immunization. (A) Flow cytometry analysis of changes in CD4 + CD25 + T cells in the hTSHR289, hTSHR290 and hTSHR410 groups after immunization. (B) Flow cytometry analysis of changes in CD8 + CD122 + T cells in the hTSHR289, hTSHR290 and hTSHR410 groups after immunization. *P<0.05 (One-way ANOVA with Tukey's post hoc test. hTSHR, human thyroid-stimulating hormone receptor.

Article Snippet: The analyte detectors were as follows: CD3 + antibody (cat. no. 565643; Becton, Dickinson and Company), CD4 + antibody (cat. no. F21004A02; Multi Sciences Biotech), CD8 + antibody (cat. no. F2100801; Multi Sciences Biotech), CD25 antibody (cat. no. E-AB-F1102C; Wuhan Elabscience Biotechnology Co., Ltd.) and CD122 antibody (cat. no. E-AB-F1029D; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Establishment of a humanized Ph + ALL xenotransplant model using anti-CD122-conditioned NOD/SCID mice and intra-bone marrow injection (IBMI) with LPCs from Ph + ALL patients. a Compared with an irradiated non-transplanted control mouse (Ctrl), the mouse transplanted with LPCs and treated with vehicle (Vehicle) exhibited significant splenomegaly at 12 weeks post-transplant. A low-magnification image of human Ph + ALL engraftment in a bone section ( left panel ), May-Giemsa staining ( middle panel ) and fluorescence in situ hybridization (FISH) analysis ( right panel ) of leukemic blasts in the BM of the recipients (Vehicle). b Flow cytometric analysis demonstrated that the BMs of the recipients were efficiently engrafted with human Ph + ALL cells with an aberrant phenotype similar to that in the donor Ph + ALL patients. c Human engraftment of the BM, spleen, liver and kidney of the recipient (Vehicle) was further confirmed by HE staining ( upper panels ) and IHC with anti-hCD19 antibody ( lower panels )

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: Injection, Irradiation, Staining, Fluorescence, In Situ Hybridization

Journal: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Effects of single drug treatment or different combinations of IM, NL and RUX on the engraftment of human Ph + ALL cells in the anti-CD122-conditioned NOD/SCID recipients transplanted with LPCs from Ph + ALL patients. a Experimental design for the in vivo experiments. LPCs sorted from newly diagnosed Ph + ALL patients (N = 6) were transplanted by IBMI into 5-week-old, sub-lethally irradiated (2.1 Gy of total body irradiation from a 60 Co source) and anti-CD122-conditioned NOD/SCID mice. From +14 days post-transplantation, the recipient mice were randomly administered with vehicle (10% NMP-90% PEG 300), IM (100 mg/kg/day), NL (75 mg/kg/day), RUX (30 mg/kg/day), IM (100 mg/kg/day) combined with RUX (30 mg/kg/day), or NL (75 mg/kg/day) combined with RUX (30 mg/kg/day) via oral gavage for 14 days. b The engraftment levels of human Ph + ALL CD45 + cells in the BMs and spleens of mice under different treatment conditions were analyzed by flow cytometry at 8 weeks (N = 6 patients, N = 3 mice per patient per treatment group) and c at 12 weeks post-transplant (N = 6 patients, N = 3 mice per patient per treatment group) or until the mice were moribund

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: In Vivo, Irradiation, Transplantation Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Cotreatment with NL and RUX exhibited the most effective anti-LPC effect. LPCs sorted from newly diagnosed Ph + ALL patients (N = 6) were transplanted by IBMI into 5-week-old, sub-lethally irradiated (2.1 Gy of total body irradiation from a 60 Co source) and anti-CD122-conditioned NOD/SCID mice. From +14 days post-transplantation, the recipient mice were randomly administered with vehicle (10% NMP-90% PEG 300), IM (100 mg/kg/day), NL (75 mg/kg/day), RUX (30 mg/kg/day), IM (100 mg/kg/day) combined with RUX (30 mg/kg/day), or NL (75 mg/kg/day) combined with RUX (30 mg/kg/day) via oral gavage for 14 days. a Representative images of splenomegaly in the mice under different treatment conditions and a control NOD/SCID mouse (Ctrl) without receiving the Ph + ALL LPCs transplantation. b Differences in human engraftment were further confirmed by HE staining ( upper panels ) and IHC with anti-hCD19 antibody labeling ( lower panels ) of the spleens in the recipient mice treated with the different drugs and the Ctrl mice. c The engraftment analysis of BCR/ABL -expressing BM cells using a TaqMan-based qRT-PCR assay in the recipient mice at 12 weeks post-transplant. d Representative western blots of phospho-CrkL, phospho-JAK2, and GAPDH in the bone marrow cells of humanized mice transplanted with Ph + ALL LPCs following treatment with vehicle, single agents or a combination of RUX and IM or NL mice, and Ctrl mice. All data from the independent experiments are presented as the mean ± SEM. Significance values: *** P < 0.0001

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: Irradiation, Transplantation Assay, Staining, Antibody Labeling, Expressing, Quantitative RT-PCR, Western Blot

Journal: Cells

Article Title: Tolerance Induced by Antigen-Loaded PLG Nanoparticles Affects the Phenotype and Trafficking of Transgenic CD4 + and CD8 + T Cells

doi: 10.3390/cells10123445

Figure Lengend Snippet: Antigen-encapsulating PLG/GP33 nanoparticles expand CD8 + Tregs in transgenic P14 mice. 3–4 female P14 mice were injected intravenously with either PLG/GP33 or a control PLG (PLG/OVA323 or PLG). Spleens and/or lymph nodes (pooled axillary, brachial, and inguinal) were harvested and counted 3 days later. Viable T cells were analyzed by flow cytometry. Statistical significance was determined by Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( A ) Expression of FoxP3, CD28, or CD122 was determined by geometric mean fluorescence intensity (MFI). ( B ) Sub-populations were determined by percentage of viable CD8 + cells that were double positive for CD122 and PD-1, Ly49, or CD38. ( C ) Female P14 mice ( n = 3) at various ages (young 4 weeks of age, adult 8 WOA, old 12 WOA) were injected intravenously with either PLG/GP33 or PLG/OVA323. Expansion of Tregs was determined by percentage of all viable CD8 + T cells that were double positive for CD122 and PD-1 by flow cytometry. Statistical significance between the groups was determined by considering three ages as three replicates.

Article Snippet: CD122 + cells were positively selected and removed from whole spleens by first staining with biotin conjugated anti-CD122 monoclonal antibody and then removing the stained cells with a biotin positive selection kit (STEMCELL Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions.

Techniques: Transgenic Assay, Injection, Control, Flow Cytometry, Expressing, Fluorescence

Journal: Cells

Article Title: Tolerance Induced by Antigen-Loaded PLG Nanoparticles Affects the Phenotype and Trafficking of Transgenic CD4 + and CD8 + T Cells

doi: 10.3390/cells10123445

Figure Lengend Snippet: Expanded CD8 + CD122 + Tregs reduce viability of CD4 + T cells. For each experiment, spleen cells from female P14 mice were harvested 3 days after intravenous injection with PLG/GP33 or control PLG. For each mouse, 5 separate ex vivo cultures were set up. CD4 + T cell viability was determined by flow cytometry. Statistical significance was determined by Student’s t -test. ns not significant, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( A ) Whole spleen cultures from PLG/GP33-treated P14 or untreated wild-type mice were incubated at 37 °C for 3 or 24 h with GP33 (1 μg/mL), anti-CD3 (1 μg/mL), or both. ( B ) Whole spleen cultures from P14 mice treated with either PLG/GP33 or control PLG (PLG/OVA323 or PLG) were incubated at 37 °C for 3 h with 1 μg/mL anti-CD3. ( C ) CD122 + cells from mice treated with PLG/GP33 were depleted by magnetic cell separation and incubated at 37 °C for 3 h with 1 μg/mL anti-CD3. Results are from three separate pooled experiments.

Article Snippet: CD122 + cells were positively selected and removed from whole spleens by first staining with biotin conjugated anti-CD122 monoclonal antibody and then removing the stained cells with a biotin positive selection kit (STEMCELL Technologies, Vancouver, BC, Canada) according to the manufacturer’s instructions.

Techniques: Injection, Control, Ex Vivo, Flow Cytometry, Incubation, Magnetic Cell Separation