Fig 1: Fandango physically interacts with a similar group of splicing proteins during oogenesis and embryogenesis.(A) Pull down assay from nuclear-enriched protein extracts using a polyclonal antibody of Prp19. Endogenous Prp19 interacts physically with Fandango and other subunits of the NTC/Prp19 complexes (ISY1 and CDC5L). Pre-immune serum was used in the control. (B) Size-exclusion chromatography of control and fand1 mutant protein extracts from 0–3 hr embryo collections using a Superose 6 10/300 column. After separation, each fraction was analyzed by Western blot. NTC/Prp19 complexes subunits (Prp19, Fandango, and ISY1) were part of a ∼600–800 kDa complex and also co-purified in a significantly larger complex (fraction 4 and 5). fand1 mutant protein extracts showed a significant reduction in levels of Fandango and ISY1 subunits and a size reduction of the Prp19-positive ∼600–800 kDa complex. (C) Western-blot analysis of total protein extracts from ovaries (left) and 0–3 hr embryos (right) from control and both fandango alleles, showed a reduction of Fandango and ISY1 protein levels in both tissues. Protein levels of Prp19 and CDC5L were not affected. α-Tubulin was used as loading control. Fandango Western blot is the same as shown in Figure 1I.DOI: http://dx.doi.org/10.7554/eLife.02181.008
Supplier Page from Abcam for Anti-ISY1 antibody