Fig. 3

PtaHMGB2/3 activates pPtaLHY2::LUC reporter in a gate-dependent manner. a, b Fluorescence microscopic images showing the subcellular localization of 35S::PtaHMGB2/3:YFP::tNOS (a) and 35S::YFP::tNOS (b) in Nicotiana benthamiana epidermal cells. White arrow indicates nuclear accumulation of PtaHMGB2/3:YFP. Scale bars are 50 μm. c PtaHMG2/3 mRNA expression levels obtained by qRT-PCR in poplar leaves samples transiently expressing 35S::amiRNA_PtaHMGB2/3::tNOS–35S::GFP::tNOS and transiently expressing the control construct 35S::GFP::tNOS. Data indicates mean of two independent experiments. d Luciferase activity detected in leaf discs when 35S::PtaHMGB2/3::tNOS–35S::GFP::tNOS is transiently expressed in pPtaLHY2::LUC genetic background under LD conditions. e Strip chart representing the distribution of luciferase values obtained at ZT1 and ZT25, for transiently transformed leaf discs of PtaHMGB2/3 overexpressing and control, respectively. Horizontal bar represents the median. This experiment was performed twice. A representative result is shown in this figure. f Luciferase activity detected in leaf discs when 35S::amiRNA_PtaHMGB2/3::tNOS–35S::GFP::tNOS is transiently expressed in pPtaLHY2::LUC genetic background under LD conditions. g Strip chart representing the distribution of luciferase values obtained at ZT1 and ZT25, for transiently transformed leaf discs of PtaHMGB2/3 knockdown and control, respectively. Horizontal bar represents the median. This experiment was performed twice. A representative result is shown in this figure. Control and transiently transformed luciferase measurements were compared with U-Mann–Whitney test at ZT1 and ZT25 (d, g). *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001