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. Author manuscript; available in PMC: 2021 Nov 19.
Published in final edited form as: Mol Cell. 2020 Nov 6;80(4):666–681.e8. doi: 10.1016/j.molcel.2020.10.014

Figure 6: ALS/FTLD-linked G156E mutation disrupts the LLPS of FUS due to a bulky amino acid substitution.

Figure 6:

(A) Structures of the different amino acids introduced at position 156. (B) Representative smFRET traces of 500 nM G156A, G156D, G156Q, G154E, and G156P FUS with 50 pM FRET-labeled pdU50 RNA. The dynamic fraction of >100 molecules was quantified and is visualized with a violin plot. (C) Wide-field images of 1 μM G156A, G156D, G156Q, G154E and G156P FUS with 1 μM U40 RNA and TEV protease at 2 h, 4 h, and 6 h. The fluorescent signal is 10 nM U40 RNA, and the scale bar is 5 μm. Droplet area was quantified by intensity thresholding and is plotted over time, and droplet fluorescence recovery after photobleaching is shown for the 4 h timepoint: G156E = green circle, G156A = yellow circle, G156D = yellow-green square, G156Q = lime diamond, G154E = gray circle, and G156P = purple circle. Significance was calculated using a two-tailed two-sample Student’s t-test with ns = not significant, * = p < 0.05, ** = p < 0.01, and *** = p < 0.001. Degrees of freedom were calculated using the Satterthwaite two-sample approximation.