Extended Data Fig. 6: Histone modification and subcellular localization of LoVIL1.
From: Epigenetic silencing of callose synthase by VIL1 promotes bud-growth transition in lily bulbs
a, LoVIL1 enhances H3K27me3 modification. Nicotiana benthamiana leaves were transiently overexpressed with 35S:eGFP-LoVIL1. Samples were collected 3 days after agroinfiltration. Nuclei were collected and subjected to immunoprecipitation before confocal imaging. Blue, green, and red fluorescence represent the 4ʹ,6-diamidino-2-phenylindole (DAPI)-stained nuclei, eGFP-LoVIL1 overexpressed nuclei, and methylated histone (H3K27me3), respectively. Non-transfected nuclei are indicated by yellow arrowheads while eGFP-LoVIL1 overexpressed nuclei are indicated by purple arrowheads. Scale bars, 10 μm. b, Quantification of methylation marks shown in panel a. The staining intensity of histone modification was compared by analyzing 10 pairs of transfected nuclei versus non-transfected nuclei in the same field of view. Four fields were analyzed using the ImageJ tool. Data are presented as mean ± s.e. of four biological replicates (two-sided student’s t-test). c, Subcellular localization of LoVIL1 in N. benthamiana leaves. 35S:eGFP-LoVIL1 was transiently overexpressed in N. benthamiana leaves. Images were captured after 3 days of agroinfiltration. mCherry fused Arabidopsis PLASTID RIBOSOMAL PROTEIN L24B (AtRPL24B; AT5G54600) was used as a nuclear marker. Fluorescence signals were collected using a fluorescence microscope with excitation wavelengths of 405 nm, 488 nm, and 561 nm. Scale bars, 20 μm. Consistent results were obtained at least in three biological repeats.
