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. Author manuscript; available in PMC: 2019 Dec 1.
Published in final edited form as: Nano Lett. 2019 Nov 4;19(11):8333–8341. doi: 10.1021/acs.nanolett.9b04147

Figure 4.

Figure 4.

Nanocapsules capture DAMPs from dying tumor cells and activate DCs: (a) Spectral counts of proteins, analyzed by LC-MS/MS, in the supernatant of B16F10 cells treated with 10 μM CFZ in serum-free medium (B16F10 supernatant), CFZ-pTA-alb (1 mg/mL NP), CFZ-pTA-alb (1 mg/mL NP) incubated in the B16F10 supernatant for 2 h at 37 °C and then rinsed. (b and c) BMDC activation following coculture with B16F10 cells pretreated with unformulated CFZ, CFZ-pTA-alb (200 nM CFZ-equivalent) or blank pTA-alb for 24 h, indicated by the expression of (b) CD40 and (c) CD86 on CD11c+ BMDCs. −: non-activated BMDCs; +: LPS-activated BMDCs. ***: p < 0.001 and ****: p < 0.0001, one-way ANOVA with Dunnett’s multiple comparison’s test versus non-activated BMDCs. (d and e) Phagocytosis of B16F10 cells by BMDCs: B16F10 cells were prelabeled with DiI, treated with unformulated CFZ, CFZ-pTA-alb (10 μM CFZ-equivalent) or blank pTA-alb, rinsed once and cocultured with BMDCs for (d) 4 h or (e) 24 h. BMDCs were identified with APC anti-mouse CD11c antibody and analyzed by flow cytometry. Phagocytosis (%): % of DiI+ CD11c+ cells in total CD11c+ cells. ***: p < 0.001 and ****: p < 0.0001, one-way ANOVA with Dunnett’s multiple comparisons test versus non-pretreated B16F10 cells.