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Fig. S5
Detailed characterization of human LRRC50. (A) Adapted image from a report by Muto et al. which describes a group of LRR containing proteins that all share localization to the centrosome. Based on the intracellular distribution of LRRC50 described in this manuscript and the presence of six Leucine Rich Repeats as well as a Coiled-Coil domain, we propose LRRC50 to be an additional member of this subgroup. (B) LRRC50 localization resembles that of the group of perichromosomal sheath proteins. A detailed description of the dynamic LRRC50 localization to chromosomes during the cell cycle is provided. Additionally, continuous association of LRRC50 with the centrosomes and localization to the midbody can be appreciated. We used Dapi (blue), α-LRRC50 ab75163 (green) and α-acetylated-α-Tubulin unless otherwise indicated. Nucleolus; in interphase cells, LRRC50 localizes to structures in the nucleus. We show these structures to be nucleoli based on co-localization with ectopically expressed mybbp1A-RFP (red), which is an established nucleolar marker. G2; cells that are in the G2 phase of the cell cycle show increased expression of LRRC50 in both nucleoli and nucleoplasm. One cell with duplicated centrosomes (using α-γ-Tubulin in red) can be observed, indicating this cell to be in the G2 phase. The neighbouring cell has no duplicated centrosomes and appears to be in interphase. Expression levels are generally lower, supported by the LRRC50 mRNA expression shown in Figure 5E and Figure S6C, and localization is confined to nucleoli. Prophase; upon completion of chromosome condensation at the G2/M transition, LRRC50 has associated with all foci of condensed chromosomes. Metaphase and Anaphase; after nuclear envelope breakdown, LRRC50 remains associated with the chromosomes throughout formation of the metaphase plane and the actual chromosome segregation during anaphase. Telophase and cytokinesis; during telophase when condensed chromosomes begin to de-condense, LRRC50 relocalizes from the perichromosomal sheath to a more diffuse and less defined chromosomal location. Followed by a temporal redistribution to the midbody.