Fig. 3: Directed screening of anionic peptides for the cytosolic delivery of chemically-modified UbcH7_86C.

a Representative images of HepG2 cells treated with 1.5 µM five anionic peptide-modified UbcH7_86C-T, respectively, in the presence of 3 µM TAT₃ for 30 minutes at 37 °C. Scale bars, 20 µm. b Quantification of the nuclear MFI of the cells in Fig. 3a. The data are presented as the mean ± standard deviation. P-values were calculated via one-way ANOVA with Tukey’s test correction. c Quantification analysis of nuclear TAMRA mean fluorescence intensity (MFI) of cells in Supplementary Fig. 6a. Data are presented as mean ± standard deviation. P-values were calculated by one-way ANOVA with Tukey’s test correction. d Quantification of the nuclear MFI of the cells in Supplementary Fig. 6c. The data are presented as the mean ± standard deviation. P-values were calculated via one-way ANOVA with Tukey’s test correction. e Images of HepG2 cells treated with 1.5 µM NLS-UbcH7_86C^T+-FITC and 3 µM TAT₃ for 30 minutes at 37 °C. The images shown are representative of independent biological replicates (n = 3). Scale bars, 20 µm. f Quantification of the fluorescence intensity along the white line shown in e. The gray box denotes the nuclear region. g CD spectra of UbcH7_86C, UbcH7_86C ⁺ and GSH-preincubated UbcH7_86C ⁺. h Confocal microscopy images of HepG2 cells treated with 1.5 µM UbcH7_86C ⁺-T or UbcH7_86C-T in the presence of TAT₃ for 30 minutes at 37 °C. The images shown are representative of independent biological replicates (n = 3). Scale bars, 20 µm. i Flow cytometry analysis of the HepG2 cells shown in Fig. 3h via detection of the MFI. “B”, “R” and “G” stand for Hoechst (blue), TAMRA (red) and FITC (green) signals, respectively. Source data are provided as a Source Data file.