Figure 3 from Phase Separation Mediates NUP98 Fusion Oncoprotein Leukemic Transformation

Abstract

<p>Mutation of multiple F and FG residues in the FG-rich IDR of NHA9 alters puncta formation in cells. <b>A,</b> Schematic of NHA9 and mutant constructs used in this study. FG motif valence is shown on the left. <b>B,</b> Representative image of live HEK293T cells expressing G-NHA9–8FA (top, green) and G-NHA9–21FGAA (bottom, green). DNA is stained with Hoechst dye (blue). <b>C–F,</b> Plots of puncta # (/10<sup>3</sup> μm<sup>3</sup>; <b>C</b>), <i>V<sub>p</sub></i> (μm<sup>3</sup>; <b>D</b>), <i>K<sub>p</sub></i> (${K_p}{\rm{\ }} = {\rm{\ }}\frac{{[ {{\rm{DP}}} ]}}{{[ {{\rm{LP}}} ]}}$ (<b>E</b>), and Δ<i>G<sub>Tr</sub></i> (kcal/mol; <b>F</b>) versus [G-NHA9 construct] for G-NHA9 (green), G-NHA9–8FA (purple), and G-NHA9–21FGAA (blue) from data represented in <b>B</b>. Data are plotted on a semi-log (<i>y</i>-axis: log<sub>10</sub>) scale in <b>C–E</b>. Refer to Supplementary Table S2 for mean values ± standard error. The pairwise <i>P</i> value between G-NHA9 versus G-NHA9–8FA and G-NHA9 versus G-NHA9–21FGAA is shown in each plot (<b>C–F</b>; <i>n</i> = 935, 683, and 865 in <b>C</b> including the cells with zero punctum and <i>n</i> = 378, 273, and 159 in <b>D–F</b> excluding the cells with zero punctum, respectively, for G-NHA9, G-NHA9–8FA, and G-NHA9–21FGAA).</p>

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