Figure 7.

Characterization of anionic liposomes (ALs).

Notes: (A) Shown is a representative, intensity-based size distribution of ALs as determined by DLS. (B) The average size, PDI, and zeta potential of ALs are shown. (C) After BMDCs were incubated overnight with a series of ALs’ concentrations, cellular viability was measured using the CCK-8 assay. ALs had no detectable effect on cross-presentation of OVA. (D) The percentage of OT-I CD8⁺ T-cells that proliferated after coculture with BMDCs pulsed with OVA and different doses of ALs. (E) Representative flow cytometric histograms of (D) are shown. The effect of ALs on the lysosomal pH of BMDCs was measured by (F) AO staining and (G) LysoSensor Green DND-189 dye staining, followed by flow cytometric analysis. The data were analyzed by one-way ANOVA, followed by a multiple comparison post-test. ***P<0.001. Data represent the mean ± SD from at least three repeated experiments with n =9 for (A), n =2–3 for (D), and n =2–4 for (F and G).

Abbreviations: ALs, anionic liposomes; ANOVA, analysis of variance; AO, acridine orange; BMDCs, bone marrow-derived dendritic cells; CCK-8, Cell Counting Kit-8; CFSE, carboxyfluorescein diacetate N-succinimidyl ester; CQ, chloroquine; DLS, dynamic light scattering; PBS, phosphate-buffered saline; PDI, polydispersity index; OVA, ovalbumin.