Fig. 2: 25-bp dsDNA substrates containing GGAA microsatellites can transfer into the homogeneous FUS-ERG condensates, inducing the hollow co-condensate formation.
From: Deciphering the molecular mechanisms of FET fusion oncoprotein–DNA hollow co-condensates
a Time course of hollow co-condensate formation of 5 μM GFP-FUS-ERG mixed with 0.6 μM AlexaFluor647-labeled 25 bp 4 × GGAA dsDNA at 0 min (i), 30 min (ii), 60 min (iii), and 90 min (iv). At the 0 min time point, we injected dsDNA. (v), (vi), and (vii) are representative events of hollow co-condensate formation and normalized intensity profiles from (i), (ii), and (iv). b (i) Time course of hollow co-condensate formation of 5 μM GFP-FUS-ERG mixed with 0.6 μM AlexaFluor647-labeled 25-bp random dsDNA at 0, 30, and 90 min. At the 0 min time point, we injected dsDNA. (ii) is the normalized intensity profile at the 90 min time point. c Boxplot of the mean intensity of dsDNA inside the condensates for GFP-FUS-ERG with 25 bp random dsDNA and 25 bp 4 × GGAA dsDNA. The total number N examined over one-time in vitro droplet experiments. For the boxplot, the red bar represents median. The bottom edge of the box represents 25th percentiles, and the top is 75th percentiles. Most extreme data points are covered by the whiskers except outliers. The ‘+’ symbol is used to represent the outliers. Statistical significance was analyzed using unpaired t test for two groups. P value: two-tailed; p value style: GP: 0.1234 (ns), 0.0332 (*), 0.0021 (**), 0.0002 (***), < 0.0001 (****). Exact P values are as follows: P < 0.0001 for all conditions: 4 × GGAA dsDNA 0 vs. 30 min; 4 × GGAA dsDNA 0-min vs. 90 min; 4 × GGAA dsDNA 90 min vs. random dsDNA 90 min; random dsDNA 0 min vs. 30 min; random dsDNA 0 vs. 90 min. Confidence level: 95%. Scale bar: 5 μm in a(i)–(iv). Scale bar: 2 μm in a(v)–(vii) and b(i). Source data are provided as a Source Data file.
