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Figure 3 | BMC Biotechnology

Figure 3

From: Intracellular delivery of peptides via association with ubiquitin or SUMO-1 coupled to protein transduction domains

Figure 3

(a) Activated lymphocytes were incubated for 4 h with 0.5 μM of FP6UG (lanes 1 and 3), FP7UG (lanes 2 and 4), FP6SG (lanes 5 and 7), FP7SG (lanes 6 and 8). After a PBS wash, cells were lysed either in protein loading buffer (SDS, lanes 3, 4, 7 and 8) or in RIPA buffer (RIPA, lanes 1, 2, 5 and 6). Immunoblot analysis of these extracts was performed with the antibody to GFP (upper panel) or to β-actin (lower panel). Signals corresponding to FPUG (U), FPSG (S) and β-actin are indicated on the right. (b) HeLa cells were transfected with 1 μg of plasmid encoding FP1UG, FP1SG, FP5UG or FP5SG. 24 h after transfection, cells were washed with PBS and lysed with RIPA buffer. Samples were loaded onto a 12%-SDS protein gel and an immunoblot was carried out using the antibody to GFP. The position of the signal corresponding to GFP (G) is indicated on the right. (c) Activated lymphocytes were incubated for 2 h with 2 μM of FP1G (top panels), FP1UG (middle panels) or FP1SG (bottom panels). Cells were then washed and observed by confocal microscopy. Visualisation was performed either with fluorescence (right panels) or with light transmission (left panels).

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