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. 2015 Aug;21(8):1404–1418. doi: 10.1261/rna.049551.115

FIGURE 2.

FIGURE 2.

HA-tagged deletion mutants of Symplekin, CPSF100, and CPSF73 can be stably expressed in Dmel-2 cells. (A) Schematics of Symplekin (top), CPSF100 (middle), and CPSF73 (bottom) deletion mutants used for in vivo binding assays are shown below each full-length protein. Each mutant is defined by the amino acids contained within the deletion, for example (1–271). The C-terminal amino acid of each mutant is written above the full-length CCC factor. The nucleotides targeted by dsRNA are defined by a solid rectangle. (B) HA-tagged full length (lanes 2,7,12) and N- and C-terminally truncated CPSF73 (lanes 3–5), CPSF100 (lanes 8–10), and Symplekin (lanes 13–18) deletion mutants were stably expressed in Dmel-2 cells. Whole-cell lysates from these strains were separated with SDS-PAGE, transferred to a membrane, and probed with an anti-HA antibody to confirm expression. Amino acids comprising the deletion mutant are shown following the protein name above the WB. Molecular weight (MW) markers for lanes 1–14 are on the left. MW markers for lanes 15–18 are on the right.