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. 2023 Mar 2;4(4):100570. doi: 10.1016/j.xplc.2023.100570

Figure 5.

Figure 5

HOS15 forms a complex with COP1 in vivo.

(A) BiFC assay for interaction between HOS15 and GI or COP1. The Venus fluorescent protein N terminus (VN) alone or fused to HOS15 (VN-HOS15) and the Venus fluorescent protein C terminus (VC) alone or fused to GI (GI-VC) and COP1 (VC-COP1) were transiently expressed in tobacco leaves, and the fluorescence was observed by confocal microscopy with an excitation wavelength of 515 nm for YFP. Scale bar corresponds to 50 μm.

(B) BiFC signal analysis with DAPI staining.

(C and D) Co-immunoprecipitation assay for HOS15 and GI interaction (C) and COP1 and HOS15 interaction (D). Transgenic plants overexpressing GI-HA(C) or COP1-GFP(D) were grown in 12D/12 L at 23°C for 12 days and harvested at the indicated ZT time points. Total protein was extracted and immunoprecipitated with α-HOS15 and α-GFP, respectively.

(E) COP1 and HOS15 interaction at low ambient temperature. Ten-day-old 35S:COP1-GFP overexpressing plants were transferred to 16°C for an additional 2 days and harvested at the indicated ZT time points. Total protein extracts were immunoprecipitated with α-GFP.