Figure 1

Fbxo41 assembles into SCF-complexes and targets to centrioles. (a) Fbxo41 assembles into SCF-complexes. HEK293T cells were transfected with the indicated constructs and subjected to immunoprecipitation with empty beads (EB), Fbxo41 or Fbxo21 antibody. Deleting (ΔF-box) or mutating (W577A) Fbxo41’s F-box ___domain prevented SCF-complex assembly. Fbxo21 was included as a positive control. Gel was cropped for clarity (full length blot available in Supplementary Fig. S5). (b) Fbxo41 is increasingly expressed in brain throughout development. Mouse brains were extracted at the indicated ages and immunoblotted with the indicated antibodies. Syntaxin1a was included as a positive control⁶². Gel was cropped for clarity (full length blot available in Supplementary Fig. S5). (c) Fbxo41 is expressed in neurons and not in astrocytes. High-density astrocyte or neuronal cultures were lysed at DIV11 and subjected to immunoblotting. βIII-tubulin was included to demonstrate the presence of primary cortical neurons. Gel was cropped for clarity (full length blot available in Supplementary Fig. S5). (d) Example of a primary hippocampal neuron fixed at DIV15 and immunostained with Fbxo41 (red), Pericentrin (green) and MAP2 (blue) antibodies. Endogenous Fbxo41 was observed in enrichments in the soma (open arrowhead). In a subset of neurons (such as this example), Fbxo41 was enriched at the pericentriolar region as indicated by Pericentrin (arrowhead). Scale bars, 5 and 2 μm. (e) Widefield fluorescence image of a primary DIV15 hippocampal rat neuron expressing Fbxo41^WT-FLAG and immunostained with antibodies for FLAG (green) and Cep135 (red). Imaging was performed with FLAG-tagged Fbxo41^WT due to antibody incompatibility. Scale bar, 1 μm. (f) Super-resolution dSTORM image of boxed region in (e). Scale bar, 500 nm.