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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment

Fig. 1

APG-115 suppresses alternative M2 macrophages polarization in vitro and increases M1 macrophages in vivo through activation of p53 pathway. a BMDMs were generated under the treatment with m-CSF for 7 days and then treated with IL-4 (20 ng/mL) to induce alternative macrophage polarization (M2) for 24 h in the absence or presence of APG-115. Cells were then harvested for detection of M2 macrophages (CD206+MHC-IIlow) by flow cytometry. b the mRNA expression levels of Arg-1 and Retnla in the above BMDMs induced by the treatment with IL-4 (20 ng/mL) with or without APG-115 were analyzed by RT-qPCR. Duplicated samples were tested. c Western blot analysis of p53, p21, c-Myc and c-Maf total proteins in BMDMs treated with IL-4 (20 ng/mL) with or without APG-115 (1 μM) for 0, 4, or 24 h, or sequentially treated with IL-4 and then APG-115 for 24 h each (24 h + 24 h). d Quantification of C. 0 (black bars), 4 (blue bars), or 24 h (green bars), or sequentially treated with each agent for 24 h (24 h + 24 h, red bars). e naïve BALB/c mice were treated with APG-115 (10 mg/kg, Q2D × 2 doses; n = 5). Two days after the last dose, spleens were collected, dissociated into single-cell suspensions, and stained with macrophage markers for flow cytometry analysis. Macrophages were defined as CD11b+ F4/80hi, and further analyzed for M1 macrophages by expression of MHC-II. Pooled data of percentages of macrophages gated on CD45+CD3 live cells (f) and percentages of M1 macrophages gated on macrophages (g) from five mice were plotted

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