Fig. 2: Half-FRAP differentiates condensates formed by LLPS from clusters formed by ICBS. | Nature Communications

Fig. 2: Half-FRAP differentiates condensates formed by LLPS from clusters formed by ICBS.

From: Detecting and quantifying liquid–liquid phase separation in living cells by model-free calibrated half-bleaching

Fig. 2

a–c Half-FRAP curves, including the normalized intensity in the bleached half (cyan) and the non-bleached half (magenta). Gray areas in the plots indicate the range of dip depths in the non-bleached half that correspond to LLPS. Half-FRAP curves for PLL-HA coacervates without magnesium chloride (a), PLL-HA coacervates with 150 mM magnesium chloride (b) and PLL undergoing ICBS (c) are shown. Top panels show representative snapshots before the bleach and 0, 5 15, 50, and 75 s after the bleach. p-values based on a one-sided Student’s t-test against the dip depths obtained for free diffusion are indicated. d Dip depths measured for half-bleached PLL-HA coacervates of different sizes. e Dip depths measured for PLL-HA coacervates that were half-bleached with different laser powers. f Dip depths measured for PLL-HA coacervates after bleaching different area fractions of them. Half-FRAP in living cells expressing DDX4-YFP (g), FUS-mCherry (h) or CD*-YFP (i). Top panels show representative snapshots before the bleach and 0, 5, 15, 50, and 75 s after the bleach. Scale bars, 2 µm. Error bands in a–c and g–i represent the standard error of the mean. p-values based on a one-sided Student’s t-test against the dip depths obtained for free diffusion are indicated in a-c and g-i. Source data are provided as a Source Data file.

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