Fig. 2: Cellular uptake of E3D2 peptide-modified UbcH786C.
a Schematic depiction of E3D2 peptide-modified UbcH786C. b Deconvoluted mass characterization of UbcH786CE3D2+. “*” corresponds to a molecular weight of 20002.1 Da, denoting 178 Da greater than the desired mass is attributed to the modified product of gluconoylated41 UbcH786C during protein expression. c Confocal microscopy images of HepG2 cells treated with 1.5 µM UbcH786CE3D2+-T or UbcH786C-T in the presence of TAT3 for 30 minutes at 37 °C. “B” and “R” stand for Hoechst (blue) and TAMRA (red) signals, respectively. The experiment was repeated three times. Scale bars, 20 µm. d Quantification of the nuclear TAMRA mean fluorescence intensity (MFI) of the cells treated in c. MFI is expressed in arbitrary units (a.u.). The results are the average of three biological replicates (n = 144 for UbcH786CE3D2+-T group and n = 157 for UbcH786C-T group) and presented as the mean ± standard deviation. P -value was calculated via two-sided t test. e Cell viability detected by a CCK-8 assay after treating HepG2 cells with 0, 1, 2, or 3 µM TAT3 for 30 minutes in serum-free DMEM. Treatment with 0.64% phenol was used as a control. The results are the average of three biological replicates The data are presented as the mean ± standard deviation. Source data are provided as a Source Data file.
