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Figure 3


Fig. 3. Rip11/FIP5 knockdown increases colocalization of Tf with TfR. Mock-treated (A-F) or Rip11 siRNA1-treated (J-L) HeLa cells were loaded for 30 minutes at 37°C with 5 µg/ml Tf-Alexa594 (B,C,E,F,H,I,K,L, red). Cells were then washed fixed and stained with anti-TfR antibodies (A,C,D,F,G,I,J,L, green). Yellow in C,F,I and L represents the overlap between Tf-Alexa594 and TfR. D-F and J-L are higher magnification images of boxed areas of mock-treated and Kif3B siRNA1-treated cells, respectively. Scale bars: 5 µm (A,G) and 1 µm (D,J). Arrowheads in D-F indicate organelles enriched in Tf-Alexa594. Arrows in D-F indicate organelles containing TfR but little Tf-Alexa594. Arrowheads in J-L point to large organelles enriched in TfR and Tf-Alexa594. Numbers in A and G give the percentage overlap between TfR and Tf-Alexa594. The data are mean ± s.d. overlap in 25 randomly chosen cells from three different experiments.





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