Expression of hereditary hemochromatosis C282Y HFE protein in HEK293 cells activates specific endoplasmic reticulum stress responses-1

<p><b>Copyright information:</b></p><p>Taken from "Expression of hereditary hemochromatosis C282Y HFE protein in HEK293 cells activates specific endoplasmic reticulum stress responses"</p><p>http://www.biomedcentral.com/1471-2121/8/30</p><p>BMC Cell Biology 2007;8():30-30.</p><p>Published online 24 Jul 2007</p><p>PMCID:PMC1947972.</p><p></p> expression plasmids and an NF-κB-luciferase reporter gene vector. As positive control, five hours post transfection cells transfected with NF-κB-luciferase reporter gene vector were stimulated with 2.5 μM thapsigargin for 24 h. Cells were incubated for 24 h in serum-complete medium. Luciferase production in cell lysates was measured by luminometery. Levels are expressed as light units (LU) per microgram of total protein (***, = 0.0009) compared to WT HFE. Assays were performed in triplicate and are representative of at least three separate experiments. () Degradation of IκB-α by C282Y HFE. HEK 293 cells were transfected with pcDNA3.1 (empty vector, EV), WT HFE, or C282Y HFE expression plasmids, and cytosolic extracts were prepared 24 h post-transfection. Ten-microgram of protein was assayed for IκB-α degradation by Western blotting (= 3). () HEK 293 cells (1 × 10) were transfected with pcDNA3.1 (empty vector, EV), WT HFE, or C282Y HFE expression plasmids and an IL-8-luciferase reporter gene vector. As positive control, five hours post transfection cells transfected with IL-8-luciferase reporter gene vector were stimulated with 2.5 μM thapsigargin for 24 h. Cells were incubated for 24 h in serum-complete medium. Luciferase production in cell lysates was measured by luminometry. Levels are expressed as light units (LU) per microgram of total protein of total protein (**, = 0.0031) compared to WT HFE. Assays were performed in triplicate and are representative of at least three separate experiments