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. 2012 May 17;33(6):617–626. doi: 10.1007/s10059-012-0080-8

Fig. 5.

Fig. 5.

Calcium effects on the interaction of AtTCTP with microtubules. (A) Microtubule cosedimentation assay from the wild-type plant Col-0. Microtubules were polymerized in the presence of 0.5 mM GTP at 37°C and harvested at the indicated times. AtTCTP and microtubules in the supernatant (Sup) and precipitate (PPT) of the reactions were detected by western blot analysis. One fifth of the supernatant samples relative to the precipitate were used for these assays. T, total crude extract. (B) AtTCTP interaction with MTs in the presence of EGTA or calcium. Microtubules were polymerized in the presence of 0.5 mM GTP at 37°C for 30 min and microtubule cosedimentation assays were performed with Col-0 total crude protein extracts after the addition of 5 mM EGTA or 5 mM calcium. Samples were harvested at the indicated times. (C, D) Quantitative analysis of the interaction between AtTCTP and MTs in the presence of 5 mM EGTA or 5 mM calcium. Each analysis was re peated three times. Error bar = SD (n = 3). Asterisks show significant differences with a P < 0.05 (t-test) vs. MT-bound AtTCTP in the presence of EGTA.