Fig. 2: Dimerisation of CPP-fused proteins enhances endocytosis and serum tolerance. | Nature Communications

Fig. 2: Dimerisation of CPP-fused proteins enhances endocytosis and serum tolerance.

From: Efficient intracellular delivery of proteins by a multifunctional chimaeric peptide in vitro and in vivo

Fig. 2: Dimerisation of CPP-fused proteins enhances endocytosis and serum tolerance.

a The MFI of T-GFP-treated HEK-293T cells (5 μM) in the indicated conditions (BSA, 50 g/L). b Schematic diagram of different GFP1–10-NLS/GFP-related recombinant proteins. The amino acid sequence of the leucine zipper is presented below. c Reduced SDS-PAGE (protein samples not boiled) and HPSEC analysis of purified T- and TL-GFP; the table shows the retention time and molecular weight of samples. d The MFI of T- and TL-GFP-treated HEK-293T cells and MFI of the T- and TL-GFP1–10-NLS-treated HEK-293T GFP11 cells in free-serum DMEM. e The MFI of HEK-293T cells treated with 5 μM T- or TL-GFP in the presence of various concentrations of FBS. f The MFI of TL-GFP-treated HEK-293T cells (5 μM) in the indicated conditions (BSA, 50 g/L). g The MFI (left panel) within total HEK-293T cells and the corresponding percentage of green fluorescence-positive cells (right panel) after treatment with TINNe-GFP- or TINNeL-GFP. Proteins were used at a concentration of 1 μM. h Representative microscopy images of the cytosolic fluorescence distribution in MA-104 cells treated with TINNe-GFP and TINNeL-GFP at the indicated concentrations. i The MFI (left panel) of total HEK-293T-GFP11 cells and the corresponding percentage of green fluorescence-positive cells (right panel) after treatment with TINNe-GFP1–10-NLS or TINNeL-GFP1-10-NLS at a concentration of 5 μM. j Representative microscope images of HEK-293T-GFP11 cells treated with TINNe-GFP1–10-NLS or TINNeL-GFP1–10-NLS at concentrations of 5 μM. k The MFI of HEK-293T GFP11 cells treated with TINNe-GFP1–10-NLS or TINNeL-GFP1-10-NLS in the presence of various concentrations of FBS. For (a (left panel), d– f (left panel), g (left panel), i (left panel) and k) the results shown represent the means ± s.e.m.; n = 3 biologically independent samples; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and NS no significant difference; two-tailed unpaired student’s t test. Relative MFI (fold increase) was obtained as above (Fig. 1). Percentages presented in the columns of (a and f) were obtained by dividing the MFI of treated cells in the given condition by the MFI of treated cells in free-serum DMEM. For (c, h and j) the data shown are representative of three independent experiments, respectively. For data, statistics, and exact P values, see Source Data File.

Back to article page