Fig. 3
CAR expression and effector activity of CAR-MΦs in vitro. A Percentages of CAR-positive RAW264.7 cells after treatment with various nanoformulations (phosphate-buffered saline (PBS), free plasmid, P/PB, P/PB/N, P/PB/N/R; P/PB contains plasmid DNA and PBAE C32-122, P/PB/N contains plasmid DNA and PBAE C32-122-NLS, and P/PB/N/R contains plasmid DNA, PBAE C32-122-NLS and PGA–RP-182 peptide. PBS and free plasmid were used as control groups). B Data are presented as a histogram and percentages. Data are shown as the mean ± SEM (n = 3). C CAR expression in BMDMs, RAW264.7 cells, GL261-H cells and 293 T cells after treatment with PCD68/PB/N/R nanoparticles or PCMV/PB/N/R nanoparticles determined by flow cytometry and D Western blotting. E Flow cytometry analysis of the M1-related marker CD80 in RAW264.7 (left) and CAR-MФ (right) cells. F LDH cytotoxicity assay of CAR-MФs. Cytotoxicity was evaluated at different effector: target cell ratios in GL261-H cells. Data are representative of three independent experiments. G Confocal images and H statistical graph of phagocytotic ability of RAW264.7 and CAR-MФ for the uptake of ErbB2 beads. Images from left to right show DAPI-stained cell nuclei (blue), CFSE-stained cells (green), ErbB2 beads (red), and merged images. Data are shown as the mean ± SEM (n = 3). Statistical significance was calculated via a two-tailed Student’s t test. **P < 0. 01; ***P < 0.001; ****P < 0.0001