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Figure 6


Fig. 6. HIF1{alpha} depletion downregulates laminin-332. (A) Real-time quantitative PCR analysis of LAMA3 mRNA expression in HaCaT cells transfected with a nonrelevant siRNA (siCtrl) (grey) or a siRNA to HIF1{alpha} (siHIF1{alpha}) (black). Q-PCR analysis was performed using transfected cells either not wounded (NW) or harvested 6 hours after wounding (W). Results were obtained from three independent experiments performed in triplicate and are expressed as normalised LAMA3 mRNA `fold increase' relative to the siCtrl-transfected non-wounded cells (siCtrl-NW). (B) Immunofluorescence detection of HIF1{alpha} (green) and laminin-332 (red) in HaCaT cells transfected with the nonrelevant siCtrl, a siRNA to HIF1{alpha} or a siRNA targeting PHD2 (that stabilised and upregulated HIF1{alpha} expression). Cells transfected with the siPHD2 siRNA were not wounded. Cells were fixed 15 hours after either scratch wounding (Wounded) or CoCl2 treatment (Co2+ treated). Nuclei were stained using DAPI. Scale bar: 40 µm.





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