Fig. 2: Virilizer acts as a scaffold between Hakai and Fl(2)d.
From: Hakai is required for stabilization of core components of the m6A mRNA methylation machinery
a Schematic representation of proteins and protein fragments used for Drosophila co-immunoprecipitation assays. b–e Co-immunoprecipitation experiments were carried out with lysates prepared from S2R+ cells transfected with Myc-GFP-tagged Virilizer (full length or fragments) and HA-tagged Hakai-long (b), Myc-GFP-tagged Hakai-long (full length or fragments) and HA-tagged Virilizer (c), Myc-GFP-tagged Virilizer (full length or fragments) and HA-tagged Fl(2)d (d), Myc-GFP-tagged Fl(2)d (full length or fragments) and HA-tagged Virilizer (e). In control lanes, S2R+ cells were transfected with Myc-GFP alone and an identical HA-containing protein. Extracts were incubated with magnetic agarose GFP binder beads and immunoblotted using anti-Myc and anti-HA antibodies (b–e), as indicated. Two percent of input was loaded. The experiment was performed in the presence of RNase A. Images shown are representative of two biological replicates. f Co-immunoprecipitation experiments were carried out with lysates prepared from S2R+ cells transfected with Myc-GFP-tagged Hakai-long and HA-tagged Fl(2)d upon control (LacZ) or vir KDs. In control lanes, S2R+ cells were transfected with Myc-GFP alone and an identical HA-containing protein. Extracts were incubated with magnetic agarose GFP binder beads and immunoblotted using anti-GFP and anti-HA antibodies. Two percent of input was loaded. g, h Schematic representing the interaction domains between Hakai, Vir and Fl(2)d derived from co-IP experiments in Drosophila (g) or HEK393T cells (h). Orange dotted lines indicate the second interaction domain between Human WTAP and VIRMA. Source data for western blots are provided as a Source Data file.
