Fig. 3: Phosphorylation of TaPIF4 associates with heat tolerance in wheat.
a Phenotypic and statistical analysis of Fielder and TaPIF4 knockout mutants in response to heat stress (42 °C for 2 days). Scale bars = 1 cm. Statistical analysis was performed with GraphPad Prism 7 (v7.00). Error bars represent standard deviation, columns labeled with different alphabet were considered significantly different (P < 0.05, one-way ANOVA analysis-Tukey comparison; n = 3 biologically independent experiments). b TaSG-D1/TaSG-D1E286K phosphorylate TaPIF4 in vitro. The phosphorylation status of TaPIF4 was examined in a Phos-tag gel assay (top panel, anti-GST). TaPIF4-P and its arrow represent the phosphorylated band of TaPIF4. TaPIF4 and its arrow represent the unphosphorylated band of TaPIF4. CIAP: calf intestinal alkaline phosphatase. The protein abundances of TaPIF4 (middle panel, anti-GST) and TaSG-D1/TaSG-D1E286K (bottom panel, anti-His) are shown in the SDS-PAGE gels. Three independent experiments were performed. c TaSG-D1 mediates the phosphorylation of TaPIF4 and its mutated form TaPIF4A in an in vitro phosphorylation assay. The phosphorylation levels of TaPIF4 and TaPIF4A were examined in a Phos-tag gel assay (top panel, anti-GST). TaPIF4-P and its arrow represent the phosphorylated band of TaPIF4. TaPIF4 and its arrow represent the unphosphorylated band of TaPIF4. CIAP: calf intestinal alkaline phosphatase. The protein abundances of TaPIF4/TaPIF4A (middle panel, anti-GST) and TaSG-D1 (bottom panel, anti-His) were monitored by SDS-PAGE. Three independent experiments were performed. Source data are provided as a Source Data file.
