FIGURE 3.

Expression of M1 3/4EGS in vivo leads to the reduction of TOP2 and SRG1 RNA. (A) Measurement of the steady-state levels of M1 3/4EGS and TOP2 RNAs in vivo. Cells transformed with empty vector pYES2.0, M1T12, or M1T28, grew in SC liquid medium for 2 d, and then diluted to OD₅₆₀ = 0.5 by SC medium containing 2% glucose (Glu) or galactose (Gal) for one more day. Cells were then collected for total RNA extraction and Northern blot analysis. The ratio for cells carrying pYES2 plasmid and grown in glucose-containing medium was chosen as 1 to calculate the relative TOP2 mRNA level in other samples. (B) The steady-state levels of 3/4EGS15 and M1 3/4EGS15 RNAs in vivo. Cells transformed with empty vector pCu426CUP1 and Cu²⁺-inducible plasmids, S15 or M1S15, grew on SC liquid medium for 2 d, and were then diluted to OD₅₆₀ = 0.5 with SC medium in the presence (+) or absence (−) of 1 mM Cu²⁺ for one more day. After that, cells were collected for total RNA extraction and Northern blot analysis. The ratio for cells carrying pCu426CUP1 plasmid and grown in the absence of Cu²⁺ medium was chosen as 1 with actin mRNA as a control to calculate the relative SRG1 RNA level in other samples.