Figure 5: Characterization of HeLa cells stably expressing a low level of His10-SUMO-2.
From: System-wide identification of wild-type SUMO-2 conjugation sites
(a) HeLa cells and HeLa cells stably expressing His10-SUMO-2 were harvested and lysed. Lysates were size-separated by SDS–PAGE, transferred to membranes, probed using a SUMO-2/3 antibody and visualized using chemiluminescence. Ponceau-S is shown as a loading control. The experiment was performed in biological duplicate. (b) HeLa cells and HeLa cells stably expressing His10-SUMO-2 were cultured on glass slides, fixed and immunostained for SUMO-2/3. Cells were investigated using confocal microscopy and immunostained SUMO-2/3 (red) and GFP (green) were visualized. Non-fused GFP is present due to expression of the His10-SUMO-2-IRES–GFP construct. The merged column is a combined display of SUMO-2/3 and GFP. Differential interference contrast (DIC, grey) scanning was performed to visualize the physical contours of the cells. Scale bars, 10 μm.
