Supplementary Figure 1: Recombinant DNMT3A-PWWP^W330R is stably expressed but has impaired H3K36me2 and H3K36me3 binding.

a, Left, Coomassie blue–stained SDS–PAGE gels with eluates from GST-PWWP protein purifications show that PWWP^WT and PWWP^W330R can be purified from bacterial lysates, in contrast to PWWP domains containing the TBRS overgrowth mutations W297del and I310N. Representative image of n = 2 independent experiments. Right, immunoblot probed with anti-GST antibody on total bacterial protein lysates (T) and purified GST-PWWP protein after elution (E) from glutathione beads, demonstrating that PWWP^W297del and PWWP^I310N are expressed but present at greatly reduced levels. b, MODified Histone Peptide Array representing 384 distinct or combinatorial histone modifications probed with recombinant PWWP proteins as indicated. Shown is an independent experimental replicate (from Fig. 2e; n = 2 independent experiments) for 10 nM PWWP protein and also after probing the array with a higher concentration (100 nM; n = 1 experiment) of PWWP^W330R protein. A Myc-tag-positive control was detected with a Myc tag antibody on the latter. Each peptide is spotted in duplicate (left and right halves of the array). Circles on the left half indicate the position of the indicated histone modification (H3K36me2 and H3K36me3) and Myc-positive control peptide on the array.