Fig. 3: YAP engages with endogenous Med1/Pol2 transcriptional condensates and drives their formation.
From: YAP charge patterning mediates signal integration through transcriptional co-condensates
A Light-sheet images of total Halo-YAP protein in nuclei with increasing YAP expression levels (left to right). Nuclear clusters (magenta circles) are only visible in nuclei with low/intermediate YAP levels. Nuclei are outlined in yellow. Images are individually min/max scaled to the YAP intensity of each nucleus. Average nuclear YAP levels are indicated. Scale bar: 5 μm. B Kymograph of total nuclear Halo-YAP protein reveals YAP clusters containing few molecules, inconsistent with one-component macromolecular condensates. The molecule number per cluster (magenta outline) was estimated from single molecule signals. Scale bar: 10 s . C 1-Cumulative Distribution Function (1-CDF) of YAP cluster intensity (expressed as molecule number per spot) quantified from total YAP light-sheet time series as shown in B. Shown are median with upper limit from 5 independent experiments. D Quantification of the YAP intensity in the dense phase (right y-axis, integral YAP cluster intensity per nucleus, norm to nuclear area) and soluble phase (left y-axis, average nuclear YAP intensity, excluding YAP clusters) as a function of average nuclear YAP concentration from images as shown in (A). Each datapoint represents one nucleus. The gray-shaded background indicates endogenous YAP levels. Shown data is pooled from N = 6 independent experiments. The graph is a zoom-in of a larger data range, see Fig. S2.E, F Simultaneous light sheet microscopy of endogenous SNAP-Med1 or Halo-Pol2 condensates (bright nuclear spots, left) and Halo-YAP or SNAP-YAP, respectively. Med1 and Pol2 condensates were identified as bright nuclear spots (left image). Kymographs (right images) show transient recruitment of YAP to Med1 and Pol2 condensates. Scale bar, 20 s. G Quantification of Med1 condensate intensity (integral per nucleus, norm to area) as a function of nuclear YAP levels from maximum projection of confocal image stacks. Shown are mean +/− SEM from N = 6 independent experiments. p values from two-sided unpaired Student’s t test.
