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. 2020 Oct 5;8(2):e000905. doi: 10.1136/jitc-2020-000905

Figure 4.

Figure 4

Depletion of macrophages impairs the antitumor efficacy of pep-20 in vivo. C57BL/6 mice were transplanted with 1×106 MC38 cells on the right flank, until the tumor volume reached 50 mm3. (A, B) Mice were treated s.c. with 2 mg/kg of pep-20 or normal saline as the negative control at the peritumoral site every day, starting on day 8 for 2 weeks and 150 µL clodronate liposomes or control liposomes were injected i.p. on days 6, 10, 14 and 18. (n=5 to 6). (C) Tumors were detected for the percentage of tumor-infiltrating CD8+ T cells in total CD45+ cells (n=3). (D, E) Cells from draining lymph nodes (D) and spleens (E) were obtained and stimulated with 20 ng/mL of PMA and 1 µM ionomycin-containing protein transport inhibitor cocktail for 4 hours. Frequencies of IFN-γ-expressing CD8+ T cells were detected by flow cytometry (n=5 to 6). Data are represented as means±SEM. Statistical significance was determined by unpaired Student’s t-test. *p<0.05, **p<0.01. IFN, interferon; i.p. intraperitoneally; n.s., no significance; PMA, phorbol 12-myristate 13-acetate; s.c., subcutaneously.