Fig. 1
Identification and affinity purification (AP) of Snf5 and Saf5: α-FLAG/M2 recognizes the FLAG tag on FZZ in the whole-cell extract (WCE) and AP experiments. a Expression analysis/AP of Snf5-FZZ. Snf5 ~ 60 kDa (18 kDa FZZ + 42 kDa Snf5). Lanes 1, 3 and 5 are untagged (−), and lanes 2, 4 and 6 are tagged (+, SNF5-FZZ) Tetrahymena strains. Lanes 1 and 2 represent WCE prepared with TCA precipitation. Lanes 3–6 were extracted with a soluble-affinity buffer. Some protein degradation is apparent. Snf5 co-purifies with Brg1 (top panel, lane 6), b expression analysis/AP of Saf5-FZZ. Saf5 ~ 83 kDa (18 kDa FZZ + 65 kDa Saf5). Lanes 1, 2, 5 and 6 are untagged (−), and lanes 3, 4, 7, 8 and 9 (positive control) are tagged (+, SAF5-FZZ) Tetrahymena strains. Saf5 co-purifies with Brg1 (top panel, lanes 7 and 8); c Snf5 and Saf5 localize to the MAC during growth. Left panels show stained nuclei, macronucleus (MAC) and micronucleus (MIC), by DAPI. Right panels show localization of the FZZ during vegetative growth