Figure 2.
Figure 2. Influence of temperature on GFP–CLIP-170 behavior. (A) GFP–CLIP-170 comets analyzed in stably expressing 3T3 cells (13 MT ends analyzed at 37°C, 19 at 32°C, and 14 at 27°C). (B) Mean fluorescence decay of GFP–CLIP-170–labeled MT ends in stably expressing 3T3 cells measured at 37°C (black; 13 ends analyzed) and at 27°C (red; 12 ends analyzed). A fit (see Materials and methods) yields a half-life of GFP–CLIP-170 fluorescence of 1.35 s at 37°C and 2.67 s at 27°C. (C) Mean fluorescence decay of GFP–CLIP-170–labeled MT ends in transiently transfected COS-7 cells at 37°C (black) and 27°C (red). See Table I for values. (D and E) Fast FRAP analysis at 27°C in MEFs transiently transfected with GFP–CLIP-170 (D) or GFP–CLIP-170XmnI (E). Fluorescent recovery was measured in the cytoplasm (blue) and on MT ends (red). For krecovery values, see Table I. Error bars indicate SEM.

Influence of temperature on GFP–CLIP-170 behavior. (A) GFP–CLIP-170 comets analyzed in stably expressing 3T3 cells (13 MT ends analyzed at 37°C, 19 at 32°C, and 14 at 27°C). (B) Mean fluorescence decay of GFP–CLIP-170–labeled MT ends in stably expressing 3T3 cells measured at 37°C (black; 13 ends analyzed) and at 27°C (red; 12 ends analyzed). A fit (see Materials and methods) yields a half-life of GFP–CLIP-170 fluorescence of 1.35 s at 37°C and 2.67 s at 27°C. (C) Mean fluorescence decay of GFP–CLIP-170–labeled MT ends in transiently transfected COS-7 cells at 37°C (black) and 27°C (red). See Table I for values. (D and E) Fast FRAP analysis at 27°C in MEFs transiently transfected with GFP–CLIP-170 (D) or GFP–CLIP-170XmnI (E). Fluorescent recovery was measured in the cytoplasm (blue) and on MT ends (red). For krecovery values, see Table I. Error bars indicate SEM.

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