Journal: Biochemical pharmacology

Article Title: The JAK1/2 inhibitor baricitinib suppresses eosinophil effector function and restricts allergen-induced airway eosinophilia.

doi: 10.1016/j.bcp.2021.114690

Figure Lengend Snippet: Fig. 2. Baricitinib and tofacitinib similarly inhibit eosinophil differentiation and JAK1/2 activation. Bone marrow cells were collected from the femurs and tibiae of three female BALB/c mice and cultured at 106/mL in media supplemented with 100 ng/mL SCF and 100 ng/mL FLT3L from day 0 to day 4. On day 4, media were replaced with media containing 10 ng/mL IL-5 supplemented with or without baricitinib (BARI, 200 nM) or tofacitinib (TOFA, 200 nM). (A) Total cells, (B) Siglec F positive cells and (C) CCR3 positive cells were enumerated by flow cytometry on the indicated days. On day 12 (D) JAK1 and (E) JAK2 phosphorylation was measured by flow cytometric intracellular staining. Data are shown as mean ± SEM from 3 independent experiments, ** P < 0.01, *** P < 0.005; (A-C) two-way ANOVA and (D and E) one- way ANOVA.

Article Snippet: On day 12, cells were processed with Fix & Perm (Nordic MUbio) and additionally stained with FITC anti-mouse phospho-JAK1 and JAK2 antibodies (both Biorbyt), evaluated by flow cytometry (BD FACSCanto II) and and analyzed by FlowJo 10.7.1 software.

Techniques: Activation Assay, Cell Culture, Flow Cytometry, Phospho-proteomics, Staining