Extended Data Fig. 4: The PS domains with non−random sequence features of FBL are required for the growth of AML cells.
a and b, FBL with PS-competent domains rescues growth defects induced by FBL deficiency in HL-60 cells (negative-selection competition assay). n = 3 biological replicates. Two-way ANOVA with Dunnett’s test. c and d, Cell cycle analysis in FBL-depleted HL-60 cells expressing ectopic FBL or mutants. n = 3 biological replicates. Two-way ANOVA with Dunnett’s test. e and f, chimeric PS competent-mutant PLD-MD fails to rescue FBL deficiency-induced growth (e) and colony formation (f) defects. PLD (prion-like domain) is another IDR from hnRNPA1 protein. g, Schematic of FBL domain organization and chimeric mutants (top). Protein disorder prediction of these sequences by IUPred3 (middle) and single amino acid charge distribution analysis along these sequences by EMBOSS (bottom). NCPR represents the net charge per residue. h, Amino acid compositional profile of sequences in g. “Polar” includes Gly, Ser, Thr, Asn, Gln, and Cys. “Aromatic” includes Phe, Tyr, Trp, and His. “Charged” includes Asp, Glu, Arg, and Lys. “Hydrophobic” includes Leu, Iso, Val, and Met. i, Quantitative summary of the area of droplets formed by FBL and its indicated mutants in AML cells (related to Fig. 4i). n = 31 (FBL, MUT-E and GAR), 33 (4KR, 4KQ, MD, H2B-MD and PLD-MD), 35 (MD-GAR) and 36 (RGG-MD) cells from 3 independent replicates. Kruskal-Wallis (Dunnett’s multiple comparisons) test. Mean ± SEM (a, b and d-f).
