Fig. 5
From: Wheat CBL-interacting protein kinase 23 positively regulates drought stress and ABA responses
TaCIPK23 interacts with TaCBL1 on the plasma membrane. a Yeast two-hybrid analysis of TaCIPK23 interaction with TaCBL1. b BiFC assay of the interaction of TaCIPK23 with TaCBL1. Fluorescence and bright field images of wheat protoplast co-transfected with constructs encoding fusion proteins TaCIPK23-YFPN and TaCBL1-YFPC. Bar = 12 μm. c In vitro pull-down assays were performed by incubating His-TaCIPK23 with GST-TaCBL1 or GST. GST-TaCBL1 was able to pull down His-TaCIPK23 fusion proteins (upper middle panel), but not GST (upper right panel) as detected by Western blot analysis with His antibodies. Coomassie Brilliant Blue (CBB) staining was used to visualize His-TaCIPK23 fusion proteins, GST and the GST-TaCBL1 fusion proteins used in each assay. d Localization of TaCIPK23 and TaCBL1 proteins in wheat protoplasts. Images were observed under a laser scanning confocal microscope. Bar = 12 μm. AtPIP1-mCherry protein was used as membrane localization marker. e Volume change of wheat protoplast transfected with GFP alone, TaCBL1-mCherry alone, TaCIPK23-GFP alone, and TaCBL1-mCherry/TaCIPK23-GFP constructs, respectively, with PEG6000 and ABA treatments. Images were observed under a laser scanning confocal microscope. Bar = 12 μm