Fig. 3: The essential function of MPP8 is independent of its chromo- and ankyrin-repeat domains but dependent on its very C-terminal region.

a Graph showing MPP8 amino acid sequence (X-axis) against the fold change reached for each sgRNA (Y-axis) used in the screen when comparing median normalized read counts from day one and day eleven. sgRNA 24 and 27 target the C-terminal ankyrin-repeat domain (ARD) while sgRNA 30 targets the N-terminal chromodomain. b Schematic representation of MPP8 proteins expressed in the rescue experiments. c Competition-based proliferation assays in mESCs stably expressing indicated Mphosph8 cDNA resistant to targeting by sgRNA 24 as well as Cas9. Cells were further transduced with the respective sgRNA. GFP is monitored over a time course of 12 days. An sgRNA targeting the mESC-specific essential gene Nanog served as positive controls while a non-targeting sgRNA (NegControl) served as negative control. The percentage of GFP+ cells is normalized to the day zero measurement and the measurement of NegControl at the respective day (n = 2 independent experiments). d Growth curves of Mpp8^mAID cell lines stably expressing MPP8 wild-type or the indicated MPP8 mutant proteins as well as OsTIR1 without or upon treatment with 500 µM IAA (mean ± s.d., n = 3 independent experiments). Source data are provided as a Source Data file.