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. 2016 Jan 22;11(1):e0146468. doi: 10.1371/journal.pone.0146468

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© 2016 Ye et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 5 — GST pull-down assay for determination of SMYD1-SRF interaction. (A) The GST control protein, GST-SRF and its deletion mutants were incubated with SMYD1 protein that was produced by in vitro translation in the presence of [³⁵S] methionine. Bound proteins were analyzed by autoradiography. (B) GST control protein, GST-SMYD1-C, GST-SMYD1-N and GST-Nkx2.5-H (positive control) were incubated with SRF protein, produced by in vitro translation in the presence of [³⁵S] methionine. Bound proteins were analyzed by autoradiography. (C) SMYD1 protein that was produced by in vitro translation in the presence of [³⁵S] methionine. SRF protein, which was produced by in vitro translation in the absence of [³⁵S] methionine. The Co-IP experiment was carried out as described in the Materials and Methods section. Anti-HA antibody (negative control) and anti-SRF antibody were used for IP, and the immunoprecipitates were examined by autoradiography. The results represent the averages ± mean from three independent experiments.