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Fig. 3. Localisation of GFP-Tm in S. pombe. (A) GFP autofluorescence of live DPM751 cells, show Cdc8 localises to the CAR during mitosis (left panel) and cytoplasmic filaments in interphase cells (right panel). (B) GFP-Cdc8 (top panel and green in bottom panel) colocalisation with actin (middle panel and red in bottom panel) was confirmed in merged images (bottom panel) of DPM751 cells stained with Rhodamine-conjugated phalloidin. (C) Real-time imaging of GFP-Cdc8 in live DPM809 cells revealed the dynamic nature of actin filaments in vivo. (D) Autofluorescence of DPM837 reveals the budding yeast tropomyosin TPM1 localises to the CAR in fission yeast cells. Rhodamine-phalloidin staining (Red) shows that GFP-Tpm1 (green) colocalises with actin (E). (F) Time-lapse imaging of DPM837 reveals GFP-Tpm1 recruits to a functional CAR. (G) Imaging of live DPM841 cells indicates smooth muscle Tm concentrate to the cell equator but fails to localise to the CAR in the presence of wild type Cdc8; but localises to the CAR in cells bearing the cdc8-110 allele (DPM924) when incubated at 36°C (H). (I) Overexpressed GFP–smooth-muscle-Tm concentrates to a single fluorescent amorphous cytoplasmic aggregate and brings about an accumulation of uninucleate elongated cells (GFP, green; phase, red). Bars, 2 µm.
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