FIG. 2.
Detection of TMG-capped RNAs by Midwestern and Northern blotting. (A) Midwestern blot. Total RNA was extracted from strain 1971 (SEN1) (lanes 1, 3, 5, and 7) and FWY1 (sen1-1) (lanes 2, 4, 6, and 8) at the times indicated following a temperature shift from 25 to 37°C. RNA was fractionated on a 9% polyacrylamide gel, transferred to nylon, and detected with anti-TMG antibodies (28). snR11 and snR31 decrease in abundance as a consequence of thermal inactivation of Sen1p. RNA species b shows a similar sen1-1-dependent decrease in abundance but has not yet been positively identified (28). (B) The blot shown in panel A was reprobed with 32P-end-labeled oligonucleotides complementary to snR11 and snR31. A probe complementary to SUF4, a glycine tRNA derived from an intronless precursor (24), was used for a loading control (not shown).