In Vivo Analysis of Ctf19 Phosphorylation, Related to Figure 2
(A) Ctf19 phosphorylation in arrested cultures (2h in arresting conditions; G1 – alpha factor; S phase – hydroxyurea; Metaphase – nocodazole and benomyl; arrows – phosphorylated Ctf19; ∗ – non-specific band). For lanes 1-3, the indicated strains were harvested during asynchronous growth (CTF19 – SMH372, ctf19-9A – SMH378). For lanes 4-7, the CTF19-3FLAG strain used in lane 2 was harvested after the indicated arrests.
(B) DNA content analysis for cultures used to produce samples 4-7 in panel B.
(C) Ctf19 phosphorylation was assessed for strains expressing wild-type CTF19 (SMH425), ctf19-9A (SMH426), or CTF19 and dbf4-9myc (SMH427) and arrested in G1. Dbf4-9myc blot is shown for pre-immunoprecipitation samples.
(D) Ctf19 phosphorylation was assessed for asynchronous cultures of the indicated strains (SMH372, SMH378, SMH401, SMH404, SMH402, and SMH403).