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Figure 2


Fig. 2. Wounding increases HIF1{alpha} protein concentration in keratinocyte cells. (A) Western blot detection of HIF1{alpha} from normal human keratinocytes (NHK) and HaCaT cell lysates at different times after scratch wounding. GAPDH was monitored as a gel-loading control. NW, not-wounded; W, wounded. (B) Immunofluorescence analysis of HIF1{alpha} in NHKs wounded using a micropipette tip (green labelling). Nuclei were stained with DAPI dye (blue). NW, not-wounded; W, 15 hours after wounding. In the upper panel, the dotted line corresponds to the wound incision and the arrows indicate the direction of migrating keratinocytes. The lower panel shows images depicting the activation of HIF1{alpha} in keratinocytes far from the wounded area of the cell monolayer. Scale bar: 20 µm. (C) HIF1{alpha} immunolabelling using peroxidase staining of paraffin slices obtained from mice epidermis harvested 1, 3, 5, 7 and 9 days (d) after injury. Arrows indicate HIF1{alpha}-positive keratinocytes. Scale bar: 60 µm.