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Figure 4 | BMC Biotechnology

Figure 4

From: Characterization of a modified ROCK2 protein that allows use of N6-ATP analogs for the identification of novel substrates

Figure 4

Phosphorylation of EF1α1 in vitro by ROCK2. A. EF1α1 (5 μg) was phosphorylated in vitro with W1161A ROCK2 as described in Methods. The samples were then separated by SDS-PAGE and transferred to a PVDF membrane. FLAG-ROCK2 was visualized by blotting with anti-FLAG M2 antibody, and EF1α1 was detected by anti-EF1α1 antibody. EF1α1 phosphorylation was visualized by phosphoimager. B. Fold increase in phosphorylation of EF1α1 by W1161A ROCK2 compared with no kinase for each of the Ser/Thr to alanine substitutions. Error bars represent the standard deviation of triplicate determinations from two independent experiments. C. EF1α1 was phosphorylated in vitro by W1161A ROCK2 in the presence of 10 nm or 1 μM Y27623. The figure is representative of two experiments.

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BMC Biotechnology

ISSN: 1472-6750

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