Supplementary Figure 5: Related to Fig. 5.

a-c, Western blot analysis of GFP or GFP-Nxf2 immunoprecipitation experiments using lysate from S2 cells transiently co-transfected with indicated FLAG-Panoramix expressing plasmids (relative amount loaded in immunoprecipitation lanes: 3x). d, Protein sequence alignment of Panoramix (308-446) where the experimentally tested residues are marked in blue. Shown below are predicted secondary structural elements and the conservation score for each position. e, Helical wheel representation of the predicted amphipathic α-helix (322-339) within Panoramix. The point mutations introduced to abolish the Nxf2 interaction are indicated in purple color. f, Western blot analysis of lysate from S2 cells transiently transfected with indicated FLAG-Panoramix expressing plasmids. Actin served as loading control. g, Box plots showing GFP intensity in S2 cells 2 days after transfection with plasmids expressing indicated fusion proteins or empty control (numbers indicate fold-change in median GFP intensity normalized to GFP-only control; box plot definition as in Fig. 3b). h, Western blot analysis of immunoprecipitation experiments (bait: stabilized degron mutant GFP-Panoramix) using lysate from S2 cells transiently co-transfected with indicated FLAG-Nxf2 expressing plasmids (relative amount loaded in immunoprecipitation lanes: 3x). i, Co-purification of His-SUMO-Panoramix helix with untagged Nxf2 UBA domain without linker by two rounds of Ni-NTA affinity purification. Source data: uncropped blot and gel images: Supplementary Data Set 1.