Lsm900 confocal microscopy

35 S:LAL and 35 S:MtLHCB1.4-GFP were used for transient transformation of Nicotiana benthamiana^{17 (link)}. Briefly, four-week-old N. benthamiana plants were infiltrated with Agrobacterium EHA105 containing 35 S:MtLHCB1.4-GFP (at an OD₆₀₀ value of 0.4) of volume 1 mL per plant or 1 mL 35 S: MtLHCB1.4-GFP (at an OD₆₀₀ value of 0.4) with 1 mL 35 S:LAL (at an OD₆₀₀ value of 0.4) per plant and incubated in the dark at 25 °C for 24 hours. Leaves with comparable BAR expression were observed at 488-nm wavelength for the GFP fluorescence using a Zeiss LSM900 confocal microscopy (Zeiss, Germany). At least five images were taken under identical conditions and the relative intensity of GFP was assessed using the ImageJ software. The RNA of leaves co-expressing LAL and MtLHCB1.4 were extracted and the RLM 5’-RACE (RNA ligase mediated rapid amplification of 5’ cDNA end) were performed with FirstChoice RLM-RACE kit (Invitrogen, U.S.A)^{22 (link)}. Three biological replicates were included, with 25 plants in each replicate.

After treating the cells with the drug at a concentration similar to the GIC₉₀ value, cells were fixed in 4% formaldehyde. Cells were then dehydrated with cold methanol in acetone (1:1). After washing extensively with phosphate-buffered saline, fixed cells were stained with anti-nucleophosmin antibody (SIGMA Aldrich, St Luis, MO, USA) and visualized with Alexa Fluor^TM 594-conjugated secondary antibody (Thermo Fisher Scientific, Waltham, MA, USA). DNA was counterstained with DAPI (DOJINDO, Kumamoto, Japan). The immunofluorescent signal was visualized using LSM900 confocal microscopy (Zeiss, Oberkochen, Germany).