Journal: FEBS Open Bio

Article Title: Differential heme release from various hemoglobin redox states and the upregulation of cellular heme oxygenase‐1

doi: 10.1002/2211-5463.12103

Figure Lengend Snippet: Hemoglobin‐mediated induction of heme oxygenase‐1 ( HO ‐1) expressions in mouse E10 cells. E10 cells were exposed to either ferrous, ferric, or ferryl Hb at equimolar concentration (100 µ m ) for 12 h for immunocytochemistry experiment (A) or 24 h for western blotting experiment (B). Laser confocal images showing expression of heme oxygenase‐1 (green, Alexa‐488) in E10 cells (A). E10 cells were also counterstained with Phalloidin Alexa‐647 (red) to show the cellular morphology. Nuclear staining was done with DAPI (blue). Cell lysates were immunoblotted and analyzed for HO ‐1 expression (B, upper panel). Equal loading was confirmed by reprobing the blots against β‐actin (B, lower panel). The ratio of average ( n = 3) band intensity of HO ‐1 with corresponding β‐actin was plotted as relative intensity (C), * P < 0.05 vs. control; # P < 0.05 vs. HbFe 3+ , ‘ t ’ test unpaired.

Article Snippet: Cells were visualized under a Zeiss LSM710 meta confocal microscope (Zeiss, Thornwood, NY, USA) after mounting with Fluoroshield mounting medium with DAPI (Abcam).

Techniques: Concentration Assay, Immunocytochemistry, Western Blot, Expressing, Staining, Control