mouse bone marrow mononuclear cell separation kit  (Beijing Solarbio Science)

Journal: Clinical & Translational Oncology

Article Title: Dipeptidyl peptidase 4 inhibitor reduces tumor-associated macrophages and enhances anti-PD-L1-mediated tumor suppression in non-small cell lung cancer

doi: 10.1007/s12094-023-03187-5

Figure Lengend Snippet: Anagliptin inhibits macrophage differentiation and M2 macrophage polarization by inhibiting late-phase extracellular signal-regulated kinase (ERK) pathway activation. a , Bone marrow-derived mononuclear cells (BMMs) were pretreated with or without anagliptin (100 μM) for 24 h and then treated with macrophage colony-stimulating factor (M-CSF) for the indicated times. The expression levels of p-ERK and ERK were determined by western blotting with the indicated antibodies. b–d , BMMs were either untreated or pretreated with anagliptin (50 µM or 100 µM) for 24 h and then differentiated for 6 days with M-CSF. On day 6, M-CSF-treated cells were treated with lipopolysaccharide (LPS) (100 ng/mL) and interferon-gamma (IFN-γ) (20 ng/mL) for 24 h or with interleukin (IL)-4 (25 ng/mL) for 24 h. The expression levels of p-ERK, ERK, P-signal transducer and activator of transcription 6 (p-STAT6), and STAT6 were determined by western blotting with the indicated antibodies. e , Schematic model illustrating the role of anagliptin in the monocyte-macrophage differentiation and M2 macrophage polarization

Article Snippet: Bone marrow mononuclear cells (BM-MNCs) were isolated from mouse femora and tibiae using a bone marrow mononuclear isolation kit (P6900, Solarbio, China).

Techniques: Activation Assay, Derivative Assay, Expressing, Western Blot