Figure 3
From: GFP-complementation assay to detect functional CPP and protein delivery into living cells
Validating SEE components by transfection.
(3a,b) SEE components are validated by co-transfection of CHO-K1 cells with hGFP1-10 and various Cargo_S11 fusion constructs. All S11 fusions complemented hGFP1-10, measured by increasing % fluorescent cells (3a) or increasing MFI (3b) in the viable single-cell population. In all experiments the “S11 only” control is a fusion of the [GSSG]x2 linker and S11 sequence and does not generate a detectable GFP signal, nor do empty vector and “GFP1-10 only” controls (% GFP-positive cells <0.65 or MFI <25). Error bars represent standard error of the mean between technical replicates; data is representative of three independent experiments. (3c) Fluorescence microscopy detects GFP complementation (FITC channel) in HCC827/hGFP1-10 cell populations transfected with complementing β-ACTIN_S11 or TRX_S11 constructs, compared to empty vector control. Cells are also counter-stained for endogenous β-Actin (TRITC) and nuclei (DAPI) before visualizing. Bar scale is 50 μm.
