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. 2015 Jul 15;10(7):e0131070. doi: 10.1371/journal.pone.0131070

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Fig 11 — (A) Mammalian IMPACT expressed in yeast precipitates Cdc28. Two different transformants of yih1Δ strain (BY4741) expressing either GST-IMPACT or GST alone from a galactose inducible promoter were grown to log phase in SGal. WCEs were prepared and equivalent amounts of protein (1 mg) were subjected to GST-pull-down assays. The precipitated complexes were analyzed by immunoblot for the indicated proteins. The input lanes (left-panel) contained 4% of the WCEs used in the assay. (B) CDK1 co-precipitates with Flag-tagged IMPACT in N2a cells. Undifferentiated mouse N2a cells were transfected with a plasmid expressing IMPACT fused to Flag or with the vector alone (pFLAG). Cell lysates were cleared with protein-A agarose and subjected to immunoprecipitation with anti-Flag antibodies (M2-Flag-Resin). All the precipitated material and 1% of the input material were subjected to immunoblot to detect Flag-IMPACT, CDK1, and GAPDH as negative control.