Supplementary Figures S1-S24 from Condensate-Promoting ENL Mutation Drives Tumorigenesis <i>In Vivo</i> Through Dynamic Regulation of Histone Modifications and Gene Expression
Authors: Liu Yiman , Li Qinglan , Song Lele , Gong Chujie , Tang Sylvia , Budinich Krista A. , Vanderbeck Ashley , Mathias Kaeli M. , Wertheim Gerald B. , Nguyen Son C. , Outen Riley , Joyce Eric F. , Maillard Ivan , Wan Liling 
Published: August 2024

<p>Supplementary Figure S1. Generation of a conditional knock-in mouse model for the Enl-T1 mutation. Supplementary Figure S2. Impact of Enl mutation on the peripheral blood and spleen. Supplementary Figure S3. Characterization of Enl-T1 allele expression and concurrent mutations in heterozygous knock-in Enl-T1 mouse model. Supplementary Figure S4. UBC-cre-ERT2/Enl flox-T1/+ mice develop aggressive acute leukemia following tamoxifen treatment. Supplementary Figure S5. Enl mutation leads to expansion of myeloid cells in mice in the leukemic phase. Supplementary Figure S6. Enl mutation leads to the decrease of B220+CD19+ B, CD4+T, CD8+ T cells in mice in the leukemic phase. Supplementary Figure S7. Impact of the Enl mutation on the bone marrow, peripheral blood, spleen, and thymus in mice in the pre-leukemic phase. Supplementary Figure S8. Enl mutation perturbs the normal hematopoietic hierarchy and leads to abnormal expansion of myeloid progenitors in mice in the leukemic phase. Supplementary Figure S9. Enl mutation does not lead to the expansion of myeloid progenitors in mice in the pre-leukemic phase. Supplementary Figure S10. Enl mutation promotes self-renewal properties of HSPCs. Supplementary Figure S11. Enl mutation-induced up- and down-regulated genes are related to distinct biological functions. Supplementary Figure S12. Enl mutation leads to a gain of myeloid differentiation signatures in HSPCs. Supplementary Figure S13. Mutant ENL-induced H3K27ac signals correlate with upregulation of development and inflammation associated transcriptional programs. Supplementary Figure S14. HSPCs gain H3K27me3 during differentiation in wildtype mice. Supplementary Figure S15. Differentiation-associated gain of H3K27me3 is impaired in Enl-mutated hematopoietic cells. Supplementary Figure S16. ENL mutants form condensate at key target genes and increase gene expression in HSPCs. Supplementary Figure S17. Condensate formation property correlates with mutant ENL’s oncogenic function in human CD34+ HSPCs. Supplementary Figure S18. Expression levels of different FLAG-ENL transgenes in LSK cells. Supplementary Figure S19. Condensate formation property correlates with mutant ENL’s oncogenic function in GMP cells. Supplementary Figure S20. Disrupting condensate formation by the H116P mutation reduces ENL-T1-induced increases in chromatin occupancy of FLAG-ENL, H3K27ac, and p300 at a subset of target genes. Supplementary Figure S21. Impact of mutant ENL on leukemia development and condensate formation. Supplementary Figure S22. Small molecule inhibition of the acetyl-binding activity of mutant ENL suppresses chromatin function and Hoxa cluster gene activation in LSK cells. Supplementary Figure S23. Small molecule inhibition of the acetyl-binding activity of mutant ENL impairs its chromatin and transcriptional function in HSPCs. Supplementary Figure S24. Small molecule inhibition of the acetyl-binding activity of mutant ENL inhibits its impact on the self-renewal property in HSPCs.</p>