Figure 3.

High-temperature cultivation of an rhIL-10 expression strain increases ER stress. (A) Reactive oxygen species (ROS) were monitored with dihydroethidium (DHE) and visualized by confocal fluorescence microscopy. (B) Cytosolic Ca²⁺ levels were monitored by staining with Rhod-3/AM and examined by flow cytometry. (C) Ubiquitin-conjugated rhIL-10 was purified from whole-cell lysates with Ni-affinity beads (cells normalized to an equivalent of 0.1 OD600 units) and examined by Western blotting using anti-Ub and an anti-His antibodies. All experiments were repeated three times and showed similar results. Scale bars: 10 μm.