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Fig. 4. Endogenous RCP and Rab11-FIP2 translocate to the plasma membrane upon PtdIns(3,4,5)P3 and PA synthesis in A431 cells. (A) Cells were unstimulated, treated with 100 ng ml–1 EGF, for 20 minutes at 37°C, or pretreated with 100 nM wortmannin before incubation with EGF. The cells were then fixed and processed for immunofluorescence with antibodies to RCP or Rab11-FIP2. (B) Cells were unstimulated, treated with 1 µM PMA for 30 minutes at 37°C or pretreated with 100 nM wortmannin followed by processing for immunofluorescence as above. (C) Cells were either treated with DMSO or 1 µM PMA for 30 minutes at 37°C before fixation. The cells were then stained for RCP (green) and ZO-1 (red). Scale bar, 10 µm.
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