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. Author manuscript; available in PMC: 2022 Apr 20.
Published in final edited form as: Sci Transl Med. 2020 Aug 12;12(556):eaaz6606. doi: 10.1126/scitranslmed.aaz6606

Figure 4. A single injection of cGAMP-MPs activates STING pathway and stimulates an immunogenic TME.

Figure 4.

A. Treatment scheme of B16F10 tumor-bearing mice receiving a single intratumoral injection of EP or cGAMP-MPs (30 μg cGAMP: 10 μg in each of cGAMP-S, PLGA-1, and PLGA-2) at day 7, or three intratumoral injections of soluble cGAMP at days 7, 11, and 15 (3×cGAMP-S, 10 μg per injection) after tumor inoculation. Tumors were isolated on day 16. B. qPCR analysis of Cxcl10 and Irf7 mRNA expression in tumors (n = 4). Data are shown as mean ± SEM. C. Western blot analysis of p-TBK1 and p-IRF3. GAPDH is used as an internal reference (n = 2). D, E, F. Percentages of infiltrating lymphocytes (D, CD8+CD3+ T cells and CD4+CD3+ T cells; E, NK1.1+CD3− NK cells), CD11b−CD11c+ dendritic cells (DCs, F), and myeloid cells (F, CD11b+F4/80+ macrophages, CD11b+F4/80−Ly6c+Ly6g+ neutrophils, CD11b+F4/80−Ly6c+Ly6g− monocytes, CD11b+Gr-1−CD200R3+ basophils, and CD11b+Gr-1−CD170+ eosinophils) in TME among all live cells (n = 4 to 5). Data are shown as mean ± SD. G. Representative flow cytometry measurements of activated DCs (CD86+CD11c+CD11b−) in tumors treated with different therapeutic combinations (n = 4 to 5). Quantitative analysis was shown on the right. BUV396 and PE represent BD Horizon Brilliant Ultraviolet 396 and phycoerythrin, respectively. Data are shown as mean ± SD. H. Representative flow cytometry measurements of M1 (CD86+CD11b+F4/80+) and M2 (CD206+ CD11b+F4/80+) macrophages in tumors treated with different therapeutic combinations. The ratio of M1/M2 macrophages was calculated and presented on the right (n = 4). Data are shown as mean ± SD. Statistical significance was calculated by one-way ANOVA or Student’s t test when comparing multiple or two groups, respectively. Data were compared with untreated group unless indicated otherwise. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.