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Fig. 3 | BMC Plant Biology

Fig. 3

From: Phosphorylation of Arabidopsis SINA2 by CDKG1 affects its ubiquitin ligase activity

Fig. 3

Interaction between CDKG1 and SINA2. a To test interactions by yeast two-hybrid assays, vectors were co-transformed into yeast AH109 strain in different combinations: 1) pGBKT7-SINA2 and pGADT7; 2) pGBKT7 and pGADT7-CDKG1; 3) pGBKT7-SINA2 and pGADT7-CDKG1. Transformants were inoculated onto SD Trp−Leu−medium and replica plated onto SD Trp−Leu−His−Ade− medium and incubated for 4 days. b In vitro pull-down assay. MBP-CDKG1 was used to pull down 6 × His labeled SINA2. The MBP tag alone was used as the negative control. Immobilized MBP and MBP-CDKG1 were detected by Coomassie brilliant blue staining. c In vivo BiFC assay. The plasmids YFPN-SINA2 and YFPC-CDKG1 were co-transformed by agroinfiltration into N. benthamiana leaves. Images were captured after 3 days using a confocal laser-scanning microscopy. Strong luminescence was localized in nuclei of epidermal cells. This signal was lost when SINA2(1–80) was used in place of intact SINA2

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