Fig. 2

The fate of PSLipos-NAC nanosystems incubated with Raw264.7 cells. (A) Flow cytometry analysis of Raw264.7 cells after 24 h incubation with PSLipos-H and PSLipos-L. The MFI value represents the amount of PSLipos captured by the cells. (B-C) Distribution of PSLipos in Raw264.7 cells. PSLipos were co-cultured with cells for 3 h before being replaced with fresh medium to maintain the culture for a period of time. The MFI values were detected at the indicated times (B). The distribution of PSLipos after 3 and 12 h was analyzed by confocal microscopy (C). Cell membranes were marked in red by DIL. The scale bar represents 10 μm. (D-E) After a 3 h co-incubation of cells with PSLipos-H-NAC or PSLipos-L-NAC, fresh culture medium was replaced and cells were further cultured. Extra- and intra-cellular NAC concentrations were tracked over time using a NAC ELISA kit. Background correction was performed for all the measurements to obtain the exogenous NAC from free NAC or PSLipos-NAC treatment by subtracting the endogenous NAC in M1 macrophages at the corresponding time point, ns represented no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001. The results represented the mean values ± SD. (n>3)