Extended Data Fig. 7: Subcellular localization of TaHST2 and physical interactions between TaHST2 and TaHSC701 or TaHSC702.
From: TaHST2 silencing shapes basal heat tolerance in allohexaploid wheat
a, Subcellular localization of TaHST2 in wheat leaf protoplasts under normal (28 °C) and HS (37 °C/7 h) condition. Before observation, the wheat protoplasts were transfected with constructs and incubated at 28°C for 16 h, followed by normal condition (28°C) or heat stress (37 °C) for 7 h. HDEL and AtRBP47 fused to mCherry were used as the marker for endoplasmic reticulum (ER) and stress granule (SG) localization, respectively. b, Bimolecular fluorescence complementation (BiFC) analysis of the interaction between TaHST2 and TaHSC701 or TaHSC702 in wheat leaf protoplasts under HS (37 °C/7 h) condition. Before observation, the wheat protoplasts were transfected with constructs and incubated at 28°C for 16 h, followed by heat stress (37 °C) for 7 h. AtRBP47 fused to mCherry was used as the marker for SG localization. c, Immunoblot analysis of the recombinant proteins obtained from the coimmunoprecipitation (Co-IP) assay. Red arrows indicate the TaHST2-GFP band. The tobacco leaves were infiltrated with Agrobacterium and incubated at 25°C for 55 h, leaf samples were then collected. In a-c, three biological replicates were performed, yielding similar results. Scale bars (a,b), 10 μm.
