spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.



Fig. 2. Changes in shape and F-actin distribution in nak1/orb3-167 cells. (A) Morphology of nak1/orb3-167 cells at 25°C (top) and 4 hours after the shift from 25°C to 35°C (bottom). (B) Morphological changes of nak1/orb3-167 cells up to 5 hours after temperature shift to 35°C. Exponentially growing cells were transferred from 25°C to 35°C and the proportions of rod-shaped, pear-shaped and round cells were determined for each time point. (C) F-Actin delocalization in nak1/orb3-167 cells. Wild-type (WT) and nak1/orb3-167 cells were grown at 25°C and samples taken before (0) and 80 minutes (80) and 300 minutes (300) after transfer to 35°C. The cells were fixed and stained with rhodamine-phalloidin for actin. (D) Changes of polar F-actin patches in wild-type and nak1/orb3-167 cells after a temperature shift from 25°C to 35°C. Cells were grown at 25°C and samples were taken before (time point 0) and after transfer to 35°C at time points indicated. The proportions of fixed and rhodamine/phalloidin-stained cells with polar F-actin patches was determined. (E,F) Repolarization of the actin cytoskeleton after a shift of nak1/orb3-167 cells from 35°C to 25°C. The nak1/orb3-167 cells were grown at 25°C, transferred to 35°C for 80 minutes (E, left) and shifted down to 25°C (E, right). Cells were fixed in formaldehyde and stained with rhodamine-phalloidin. (F) Samples were taken of nak1/orb3-167 cells up to 80 minutes after transfer from 35°C to 25°C (–HU, closed circles). The nak1/orb3-167 cells were blocked in 15 mM hydroxyurea for 3 hours at 25°C, then shifted to 35°C for 80 minutes and shifted back to 25°C to follow actin relocalization in blocked cells (+HU, open circles). At the time points indicated, the cells were fixed and stained with rhodamine-phalloidin. The proportions of cells with polarized F-actin patches at the cell tips. 200 cells were counted for each time point and experiments were performed three times. Bars, 10 µm.





Right arrow Return to article