Fig. 5: VIN3 and VRN5 assemblies exhibit a two-molecule spacing in their stoichiometry distributions.
From: SlimVar for rapid in vivo single-molecule tracking of chromatin regulators in plants
The number of labelled molecules in each assembly (stoichiometry) shows consistent peak-to-peak spacing via periodicity analysis of a VRN5-YFP and b VIN3-GFP across different vernalisation timepoints: NV not vernalised (yellow), V2W two weeks of cold (ochre/light green), V6W six weeks of cold (orange/dark green), V6WT14 six weeks of cold followed by two weeks of warm conditions (red). A kernel width (curve smoothing parameter) of 0.6 molecules was used corresponding to the standard deviation in the observed intensity of a single molecule at the sifting signal-to-noise threshold. Insets: Periodicity analysis - the number of molecules in this subunit can be estimated from the most common spacing between neighbouring peaks in each stoichiometry distribution. The threshold above which a null (aperiodic) distribution can be rejected is the 95th percentile fraction of intervals (grey trace) output from simulated random stoichiometry (Methods). The most common interval is given by the modal kernel density estimate ± s.e.m. above the null threshold (VIN3-GFP: V2W, 1.9 ± 0.3; V6W, 2.2 ± 0.3. VRN5-YFP: NV, 1.9 ± 0.4; V2W, 2.2 ± 0.4; V6W, 2.0 ± 0.3; V6W + T14, 2.0 ± 0.4). The periodic unit in each of these cases is consistent only with an assembly subunit of 2 molecules of either VIN3-GFP or VRN5-YFP. Source data are provided as a Source Data file.
