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. Author manuscript; available in PMC: 2023 Nov 3.
Published in final edited form as: Mol Cell. 2022 Oct 21;82(21):4080–4098.e12. doi: 10.1016/j.molcel.2022.09.034

Figure 5. Multiple regions of ENL mutants are required for condensate formation and function.

Figure 5.

(A) Schematic showing different ENL variants and their ability to form puncta under overexpression (O/E) and form puncta and exhibit gain-of-function (GOF) on chromatin at endogenous levels.

(B-D) Representative images (B) and fraction of in-puncta fluorescence intensity quantification (C and D) of HEK293 cells transiently transfected with indicated mCherry-ENL variants. B, all cells shown had similar mCherry-ENL intensities. C and D, each dot indicates one cell (n > 20).

(E and J) Flag IF staining in HEK293 cells expressing endogenous levels of indicated Flag-ENL. J, the yellow arrows show smaller nuclear puncta formed by T2(ΔIDR2).

(F, G, K, L) Percentage of nuclei with and without Flag-ENL puncta (F and K) and the number of Flag-ENL puncta in each nucleus (G and L) in HEK293 cells. Each dot indicates one cell (n at the range of 30~60). Center lines indicate median and box limits are set to the 25th and 75th percentiles. (H, I, M, N) ChIP-qPCR analysis of Flag-ENL occupancy at HOXA locus (H and M) and mRNA expression (normalized to GAPDH) of HOXA genes (I and N) in HEK293 cells with endogenous levels of indicated Flag-ENL. Data shown are Mean ± S.D..

F and K, Chi-square test. G-I, L-N: Two-tailed unpaired Student’s t-test. *** P < 0.001, ** P < 0.01, *P < 0.05.

See also Figure S5.