antibody pe-conjugated anti-mouse cd25 (pc61.5)  (Thermo Fisher)

Journal: eLife

Article Title: Thymic dendritic cell-derived IL-27p28 promotes the establishment of functional bias against IFN-γ production in newly generated CD4 + T cells through STAT1-related epigenetic mechanisms

doi: 10.7554/eLife.96868

Figure Lengend Snippet: ( A ) CD4SP (GFP + CD4 + CD8 - CD44 lo ) thymocytes, CD4 + RTEs (GFP + CD4 + CD8 - CD25 - CD44 lo ) and CD4 + naive (GFP - CD4 + CD8 - CD25 - CD44 lo ) T cells were sorted from 6- to 8-week-old Cd11c-p28 f/f mice and WT littermates, stimulated with plate coated anti-CD3 (2 µg/mL) and soluble anti-CD28 (1 µg/mL) for 12 hr. mRNA levels of Ifng, Il4, and Il2 were determined by qPCR. Data: mean ± SD (n=4, duplicates). ( B ) Sorted cells were cultured under Th0 conditions for 3 days. The frequency of IFN-γ-producing CD4 + T cells were measured by intracellular staining. Representative dot plots (left) and statistical data (right, mean ± SD, n=3). ( C ) Supernatants from 3-day cultures were analyzed for IFN-γ and IL-4 by ELISA. Data: mean ± SD (n=3). ( D ) mRNA levels of Tbx21 and Gata3 in sorted cells were determined by qPCR. Data: mean ± SD (n=4, duplicates). ( E–F ) T-bet protein levels were assessed by western blot ( E ) and flow cytometry ( F ) after 3-day culture. Data: mean ± SD (n=3). ( G ) Freshly sorted cells were lysed in Trizol, and Ifng and Tbx21 mRNA levels were determined by qPCR. Data: mean ± SD (n=4, duplicates). Statistical differences: * p <0.05, ** p <0.01, *** p <0.001 (Student’s t -test). Figure 1—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 1—source data 2. Original files for western blot analysis displayed in .

Article Snippet: Antibody , PE- conjugated anti-mouse CD25 (PC61.5) , eBioscience , Cat#: 12-0251-82 RRID: AB_465607 , FACS (5 μL per test).

Techniques: Cell Culture, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Interferon-γ-stimulated antigen-presenting cancer-associated fibroblasts hinder neoadjuvant chemoimmunotherapy efficacy in lung cancer

doi: 10.1016/j.xcrm.2025.102017

Figure Lengend Snippet:

Article Snippet: CD25 Monoclonal Antibody (PC61.5), PE-Cyanine7 , Thermo Fisher Scientific , Cat# 25-0251-81; RRID: AB_469607.

Techniques: Recombinant, Staining, Purification, In Vivo, Control, Protease Inhibitor, Activation Assay, Membrane, Modification, Bicinchoninic Acid Protein Assay, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Cell Isolation, Sequencing, Negative Control, shRNA, Software

Journal: Frontiers in Oncology

Article Title: DSP-0509, a TLR7 agonist, exerted synergistic anti-tumor immunity combined with various immune therapies through modulating diverse immune cells in cancer microenvironment

doi: 10.3389/fonc.2024.1410373

Figure Lengend Snippet: DSP-0509 reduced Treg function and differentiation, and when combined with IDO1 inhibitor, enhanced anti-tumor activity. (A) Representative histogram of CellTrace™ Violet (CTV) fluorescence. CD4 Teff cells were co-cultured with Tregs for 3 days. CD4 Teff cell proliferation was evaluated by the ratio of CTV fluorescence intensity. (B) The proliferation index was calculated based on CTV fluorescence intensity. *P < 0.01 two-tailed Student t test vs control. Values are the mean ± S.E.M. of each group, n = 3/group. (C) Tregs were differentiated from CD4 + cells in the presence of TGFβ and CD11c + DCs. Tregs were detected as CD4 + CD25 + Foxp3 + cells. *P < 0.05 for the τ-test. (D) Curve of HM-1 tumor growth. DSP-0509 5 mg/kg i.v. was administered once weekly and Epacadostat 100 mg/kg p.o. was administered once daily. *P < 0.05 for the Dunnett test. n = 6/group and the values are the mean and S.E.M. (E) TILs were isolated from HM-1 tumor at day 21. Tregs and CD8 + cells were analyzed by flow cytometry. *P < 0.05 for the Wilcoxon rank sum test.

Article Snippet: Flow cytometry analysis was carried out using the following primary antibodies: PE-CD11b (BD Biosciences, M1/70), FITC-Gr-1 (BD Biosciences, RB6-8C5), FITC-CD8a (BD 53-6.7), FITC-CD4 (eBioscience, Inc., RM4-5), PE-CD25 (eBioscience, Inc., PC61.5), APC-CD45RB (eBioscience, Inc., FJK-16s), PE-Cy7-CD11b (BD Biosciences, M1/70), and PE-CD86 (BD Pharmingen, GL1), and the following viability dyes: Fixable Viability Stain 450 (BD Horizon) and Fixable Near-IR Dead Cell Stain (Invitrogen).

Techniques: Activity Assay, Fluorescence, Cell Culture, Two Tailed Test, Control, Isolation, Flow Cytometry

Journal: eLife

Article Title: Sustained store-operated calcium entry utilizing activated chromatin state leads to instability in iTregs

doi: 10.7554/eLife.88874

Figure Lengend Snippet:

Article Snippet: Antibody , CD25 Monoclonal Antibody (PC61.5), PE, rat monoclonal , eBioscience , 12-0251-82 , 1:500.

Techniques: Purification, Produced, Recombinant, Staining, Cell Isolation, Selection, Extraction, Enzyme-linked Immunosorbent Assay, Software