FIG. 3.
Efficient expression of G1 cyclins requires Met30. (A) α-Factor arrest and release were as described in the legend to Fig. 2A. Samples were analyzed by immunoblotting: wild-type cells and met30-6 mutants expressing both Cln2-HA3 and Cln3-18myc were used to detect Cln2 and Cln3; strains expressing both Swi4-6myc and Swi6-HA3 for detection of Swi4 and Swi6. Cdc28 was detected with anti-PSTAIRE antibodies as a loading control. (B) Total RNA was analyzed by Northern blotting with radiolabeled probes to detect CLN1 and CLN2, respectively. (C) Strains as indicated were replica plated to plates lacking uracil and plates containing 5-fluoro-orotic acid and incubated at 25°C.