Fig. 4.

Glioblastoma cells have differential dependency to SET1/MLL family of transcription factors interacting with ASH2L. A Scheme of tandem mass spectrometry experiments performed on glioblastoma cells. B, C Proteomic analyses of GFP-tagged ASH2L in U373 (B) and U87MG (C) cells. Volcano plots of significant interactors of GFP-ASH2L isolated from nuclear extracts are shown. Stoichiometry plots of bound SET1/MLL family members (MLL family specific: yellow, SETD1A/B specific: red, MLL & SETD1A/B common members: blue) are depicted. All interactors are normalized to the GFP-ASH2L bait. Results shown represent Intensity Based Absolute Quantification with standard deviations. D Distribution of ASH2L common interactors in terms of subcellular localization in U373 and U87MG cells. E qRT-PCR analysis for mRNA levels upon siRNA mediated knockdown of WDR5 and KMT2A genes in U373 and U87MG cells. F Representative images of long-term clonogenic assay upon siRNA mediated knockdown of WDR5 and KMT2A genes and statistical analysis. Quantification of colonies was performed by ImageJ software. P values determined by two-tailed Student’s t-test *P < 0.05, **P < 0.01, ***P < 0.001