Figure 1

SETD2 loss of function mutations in the index MCL case (MCL1). (a) Sanger sequencing chromatograms with the frameshift (top) and nonsense (bottom) mutations identified by whole-exome sequencing. (b) Localization of the mutations with respect to the key functional domains of the SETD2 protein. The SRI domain is necessary for histone H3 lysine 36 trimethylation (H3K36Me3) and mediates SETD2 interaction with the phosphorylated C-terminal domain of the RNA polymerase II large subunit (RNA pol II) and with the heterogeneous nuclear ribonucleoprotein-L (HnRNP), thus coupling H3K36Me3 with transcription elongation and splicing. (c) from top to bottom: western blotting (WB) showing the truncated SETD2 (tSETD2) protein as compared to full-length SETD2 detectable in a pool of proteins from mononuclear cells of healthy donors; co-immunoprecipitation experiments performed by using: an anti-RNA pol II, an anti-hnRNP and an anti-histone H3, respectively, to isolate the proteins of interest and then an anti-SETD2 as primary antibody to label the PVDF membrane on which the immunoprecipitates were transferred; WB for H3K36Me3. Histone H3 and actin were used as loading controls.