Figure 5. UTY is a weaker tumor suppressor with altered condensate properties.

a. Growth curves of THP-1 cells electroporated with indicated constructs, shown as mean ± SD. n = 4 independent experiments. Statistical analyses for the last time point.

b. Turbidity, as shown by pictures and OD600, of 30 μM mEGFP, mEGFP-UTX cIDR, or-UTY cIDR. OD600 is presented as mean ± SD. n = 3 independent experiments.

c, d. Representative fluorescence images of 293T cells transfected with UTX or UTX-UTY_IDR fused to EGFP (c). d. Partition coefficient for the top 3 brightest GFP foci in each randomly selected cell. n = 10 cells for UTX, and 17 cells for UTX-UTY_IDR.

e. FRAP of 50 μM mEGFP-UTX cIDR or -UTY cIDR, at early (5’-40’) and late (120’-160’) times in condensation. Left, representative images of early droplets at indicated time points (photobleaching at 0). Right, recovery curves as mean ± SD. n = 5 UTX droplets and 6 UTY droplets. Scale bar, 2 μm.

f. Left, scatter plot of mEGFP-UTX or UTY cIDR diffusion coefficients in droplets in vitro by Line RICS assays, over time in condensation. Right, bar plot for mean ± SD of the diffusion coefficients at early (< or = 100 min) or late (> 170 min) times. n = 8 UTX droplets and 9 UTY droplets.

g. FRAP of transfected UTX or UTX-UTY_IDR fused to EGFP in 293T cells. Recovery curves as mean ± SD. Statistics between the two samples at different time points. The half time of recovery is shown as mean ± SD. n = 8 cells for UTX, and 9 cells for UTX-UTY_IDR.

h. A diagram showing the impact of condensation and the different material properties of the UTX WT, cancer mutant, and UTY on the activities in tumor suppression.

*P<0.05, **P<0.01, by two-sided t-test for all.