Casein kinase TbCK1.2 regulates division of kinetoplast DNA, and movement of basal bodies in the African trypanosome
Fig 6
Intracellular location of TbCK1.2: Identification of candidate TbCK1.2-pathway proteins.
(A) Trypanosomes expressing TbCK1.2 tagged at its N-terminus with V5 epitope were fixed with methanol and probed with anti-V5 antibody. Anti-centrin antibody 20H5 was used to stain basal bodies and bilobes, and DAPI was used to stain DNA. Control images show a V5-TbCK1.2 tagged line without addition of primary antibodies. Arrowheads indicate flagellum (F), basal bodies (BB), bilobes (BL), nuclei, (N), and kinetoplasts (K). Scale bar = 5 μm. (B) Following knockdown (24 h) of TbCK1.2, SILAC phosphoproteomics was used to identify candidate TbCK1.2-pathway proteins. The abundance ratio (H/L) of identified phosphopeptides is plotted as a function of their posterior error probability (PEP) value. Only peptides with a PEP score of 5 x 10−2 (5% chance of error), or lower, are presented. Grey area represents phosphopeptides with a 2-fold or greater increase in abundance. Blue shading indicates phosphopeptides that decreased in abundance 2-fold or greater.