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. Author manuscript; available in PMC: 2010 Sep 27.
Published in final edited form as: Mol Cell. 2009 Feb 13;33(3):365–376. doi: 10.1016/j.molcel.2008.12.028

Figure 4. WDR33 is a bona fide component of the CPSF complex.

Figure 4

(A and B) Chacterization of the immuopurified CPSF73 complex. Proteins from Flag-IP of NE from HEK293 or the CPSF73-3Flag cell line were resolved by SDS-PAGE and stained with silver (A), or analyzed by Western blotting using specific antibodies (B). Protein bands in (A) whose identities were confirmed by western are labeled. (C) Immunopurified CPSF complexes were analyzed by gel filtration using a Superdex-200 column. Proteins from input and each fraction were analyzed by SDS-PAGE and silver staining (top panel) and western (bottom panel). WDR33 is marked by a arrow. (D) Western analyses of the mock-treated (mock) or WDR33-depleted (ΔWDR33) NE. Intensities of individual bands were quantified using the Odyssey Infrared scanner (Li-Cor). (E) WDR33 is essential for 3′ processing. Mock, ΔWDR33 NE alone, or ΔWDR33 supplemented with immunopurified CPSF were used in standard cleavage assays. RNAs were resolved and visualized as described above.