Fig. 5. Translocation of GFP-Citron-K to the cortex of interphase cells by expression of dominant active Rho. (A) Effects of expression of dominant active Rho on localization of Citron-K. Myc-Citron-K and GFP-Val14RhoA were co-expressed in HeLa cells. Citron-K (red) and Val14RhoA (green) are colocalized in band-like structures at the bottom of cells (bottom slice), whereas in the middle of the cell, Citron-K is present as an aggregate without colocalization with Val14Rho (the middle slice, arrowheads). Band-like structure of Citron-K and Val14RhoA at the bottom consists of small patches. (B) Vertical views of GFP-Citron-K localization in HeLa cells co-expressing Myc-Val14RhoA and GFP-Citron-K. Note that expression of Val14RhoA (red) disintegrates cytoplasmic aggregates of GFP-Citron-K (green) and translocates it to the cell cortex to form band-like structures. Signals of myc-Val14RhoA and GFP-Citron-K appear not to colocalize completely because the fluorescence of Texas Red-Myc-Val14RhoA is not so strong as GFP-Citron-K, and only a small portion of expressed Rho colocalize with Citron-K. (C) Effects of expression of GFP-tagged wild-type RhoA (the left pair of panels) and GFP-tagged Asn19RhoA (the right-hand pair of panels) on localization of Myc-tagged Citron-K. Myc-Citron-K (red) remains as aggregates when wild-type or dominant negative RhoA (green) is co-expressed. (D) Effects of Rac expression on Citron-K localization. GFP-Citron-K (green) and either Myc-tagged Val12Rac1 (red) (the left pair of panels) or FLAG-tagged Asn17Rac1 (red) (the right pair of panels) were co-expressed in HeLa cells. Note that either expression did not affect the cytoplasmic aggregates of GFP-Citron-K. (E) Effects of Cdc42 expression. GFP-Citron-K (green) and either Myc-tagged Val12Cdc42 (red) (the left pair of panels) or Myc-tagged Asn17Cdc42 (red) (the right pair of panels) were co-expressed in HeLa cells. Note that either expression did not affect the cytoplasmic aggregates of GFP-Citron-K. C, D and E all show the bottom slices of interphase cells. Bars, 10 µm.
