Fig. 2: The killswitch arrests the dynamics of endogenous cellular condensates.
From: Probing condensate microenvironments with a micropeptide killswitch
a, Schematic of the nanobody–killswitch protein constructs. b, Schematic of the nanobody-based recruitment system in live cells. c, Example of the nucleus-highlighting strategy. The nuclear contour of mCherry (mCh)-expressing cells is highlighted with a magenta dashed line. The nuclear contour of untransfected (untr.) or mCherry− cells is highlighted with a white dashed line throughout the figure. The experiment was repeated independently three times with similar results. Scale bar, 10 µm. d, Live-cell fluorescence microscopy images of cells expressing GFP–NPM1, GFP–TCOF1, SRRM2–GFP and GFP–HP1α transfected with the indicated GFP-nb or GFP-nb–KS constructs. The GFP in each cell line is knocked into the endogenous gene locus. Scale bars, 10 µm. e, FRAP analysis of the GFP in the condensates shown in d. Data are mean ± s.d. n values are the same as described in f. f, Control quantification of GFP intensity in the bleached condensates. Data are mean ± s.d. One condensate for each cell was bleached, and one dot represents the value for one condensate. For the NPM1 plot, n = 10 cells for all cases, except for 2×KS (n = 12 cells), from one experiment. For the TCOF1 plot, n = 10 cells for all cases from two biologically independent experiments. For the SRRM2 plot, n = 10 cells for all cases, except for 2×KS (n = 9), from one experiment. For the HP1α plot, n = 19 (untransfected), 10 (Nb and 2×KSF-to-G), 12 (KS) and 13 (2×KS) cells from one experiment. P values were calculated using one-way ANOVA followed by Tukey’s post hoc test versus untransfected cells or mCherry alone; exact values are provided in the ‘Statistics and reproducibility’ section.
