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. 2003 Nov;23(21):7909–7919. doi: 10.1128/MCB.23.21.7909-7919.2003

FIG. 4.

FIG. 4.

Effect of band II RNAi on the ligases and other editing proteins. Cultures of two clonal band II RNA cell lines and 29.13 control cells were induced or noninduced, and on day 5 small-scale extracts were prepared in parallel. (A) They were assayed by adenylylation (following deadenylylation, to detect all ligases). The right two lanes of panels A and B both show a partly purified trypanosome extract fraction (after Q-Sepharose) (32), assayed without or with deadenylylation (deAn), either by adenylylation (A) or Western blotting for band V (B). In this preparation, band V is very highly preadenylylated and thus is virtually not detected unless first deadenylylated. (B and C) Western blotting for band V protein and LipDH control protein, respectively. (D) Northern blotting for band V sequences. (E and F) Western blotting for band III and band IV proteins, respectively. (G) Similar extracts from separate clones of band II RNA cells and control (29.13) cells, enriched by velocity centrifugation, were assayed by Western blotting for band I, VI, and VII proteins.