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Fig. 8. Clathrin localization on the plasma membrane and TGN is altered in cells transfected with Hsp70(K71E). (A) TIR-FM images of GFP-clathrin in cells that were cotransfected with GFP-clathrin and either Hsp70 (a) or Hsp70(K71E) (b). (B) Confocal images of cells transfected with Hsp70(K71E) and immunostained with anti-flag (a) and anti-clathrin (b) antibodies. (C) The exchange of cytosolic clathrin clusters measured in cells cotransfected with GFP-clathrin and Hsp70(K71E). Data were collected from photobleaching 30 cells. One-third of the cells recovered with a half-life of 20 seconds (solid triangles), one-third showed no significant recovery (open triangles) and the remainder of the cells recovered with a time course between these extremes. The dashed line is the time course of recovery of the cytosolic clathrin measured in the GAK-depleted cells in Fig. 6B.





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