Figure 5.

Efficient CNS Cargo Protein Delivery by dNP2

(A) The dTomato, dNP2-dTomato, and 11R-dTomato constructs. (B) SDS-PAGE analysis of purified proteins. (C) Naive (CD4⁺CD25⁻CD62L^highCD44^low) T cells were incubated with 10 μM dTomato, dNP2-dTomato, and 11R-dTomato for 1 h, and cells were analyzed by flow cytometry. (D) Μean fluorescence intensity (MFI) was analyzed in naive CD4⁺ T cells treated with various concentrations (1, 2, 5, or 10 μM) of dTomato, dNP2-dTomato, and 11R-dTomato for 1 h. (E) Eight-week-old female C57BL/6 mice were intravenously injected with 5 mg of dTomato, dNP2-dTomato, and 11R-dTomato. After 2 h, spinal cord and brain tissues were harvested and prepared as frozen slides. The nucleus was stained with Hoechst and fluorescence staining and was observed via fluorescence microscopy. Data are presented as mean ± SEM of three independent experiments (n = 3); statistical analysis by two-tailed Student’s t test. **p < 0.01, ***p < 0.001 versus PBS; ⁺⁺⁺p < 0.001 versus 11R-dTomato. NS, not significant.