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Fig. 2 | BMC Plant Biology

Fig. 2

From: Kinase-Associated Phosphoisoform Assay: a novel candidate-based method to detect specific kinase-substrate phosphorylation interactions in vivo

Fig. 2

Detection of phosphoisoform distribution of transiently expressed GFP variants by cIEF-immunoassay. a-c Electropherograms of various GFP-fusion proteins and their isoform distributions in cIEF-immunoassay. Expressed proteins and treatments are indicated for each sample. a Unmodified GFP is present in one major isoform and is not phosphorylated by MPK6. Top: control (single GFP construct transformation), middle: GFP co-expressed with MPK6, bottom: GFP co-expressed with flg22-activated MPK6. b Isoform distribution of GFP:ACS-C (the C-terminal domain of ACS6 fused to the C terminus of GFP). Top: control (single GFP:ACS-C construct transformation), middle: GFP:ACS-C co-expressed with MPK6, bottom: GFP:ACS-C co-expressed with flg22-activated MPK6. Asterisks indicate acidic isoforms specifically accumulating in the presence of activated MPK6. c Isoform distribution of a GFP:ACS-C variant, which is nonphosphorylatable by MAPKs (GFP:ACS-C-AAA). Top: control (single GFP:ACS-C-AAA construct transformation), middle: GFP:ACS-C-AAA co-expressed with MPK6, bottom: GFP:ACS-C-AAA co-expressed with flg22-activated MPK6. d Conventional SDS-PAGE immunoblot of transiently expressed GFP variants. Arrowhead indicates a band, which specifically accumulates in the presence of activated MPK6. Negative control (neg. cont.) denotes a protoplast sample not transfected with GFP

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BMC Plant Biology

ISSN: 1471-2229

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