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. 2003 Nov 10;163(3):489–502. doi: 10.1083/jcb.200303017

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Copyright © 2003, The Rockefeller University Press

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Figure 4. — Nuclear export of PLZF follows internalization of HB-EGF-C. (A and B) Time-lapse imaging of CFP-PLZF and HB-EGF–YFP in living cells. Cells cotransfected with CFP-PLZF and HB-EGF–YFP expression vectors were cultured for 24 h before observation, and then the culture medium was replaced with medium containing 100 nM TPA. Shown are representative serial images of transfected HT1080/HB-EGF (A) and HT1080/ΔMP-ADAM12/HB-EGF (B) cells taken every 15 min up to 90 min after TPA stimulation. Note that translocation of HB-EGF–YFP preceded nuclear export of CFP-PLZF in HT1080/HB-EGF cells. Changes in subcellular localization of the CFP and YFP fusion proteins were not observed after TPA treatment of HT1080/ΔMP-ADAM12/HB-EGF cells. (C) Time course–dependent changes in interaction of HB-EGF-C with CFP-PLZF. The HB-EGF-C–PLZF interaction was investigated by immunoprecipitation using an anti-HB-EGF-C antibody in HT1080/HB-EGF and HT1080/ΔMP-ADAM12/HB-EGF cells. The band of CFP-PLZF coimmunoprecipitated with HB-EGF-C (top) was evident at 15 min and the later after TPA treatment in HT1080/HB-EGF but not HT1080/ΔMP-ADAM12/HB-EGF cells, despite same expression of CFP-PLZF in cells (bottom). Bars, 10 μm.