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. 2004 Sep 9;23(21):4232–4242. doi: 10.1038/sj.emboj.7600326

Figure 3.

Figure 3

Co-purification of TFIIS with components of SAGA in cell-free extracts. (A) Co-purification of TFIIS∷3HA with immunoprecipitated Spt8∷13Myc. Strains YPH499 (MED13+) and YBV39 (med13Δ) were transformed with plasmids encoding the Spt8∷13Myc fusion (pVV225) and/or the TFIIS∷3HA fusion (pVV227). Protein extraction, immunopurification and detection were as for Figure 2A. (B) Co-purification of Spt8∷3HA with immunoprecipitated TFIIS∷13Myc. Strain YPH499 was transformed with plasmids encoding the TFIIS∷13Myc fusion (pVV234) and/or the Spt8∷3HA fusion (pVV230). Protein extraction, immunopurification and detection were as in Figure 2A. (C) Co-purification of Gcn5∷13Myc and Spt7∷13Myc with immunoprecipitated TFIIS∷3HA. Strain YPH499 was transformed with plasmids encoding the TFIIS∷3HA fusion (pVV228) and/or the Gcn5∷13Myc (pVV231) or the Spt7∷13Myc (pVV232) fusions. Protein extraction, immunopurification and detection were as for Figure 2C. (D) Co-purification of TFIIS∷3HA with immunoprecipitated Gcn5∷13Myc and Spt7∷13Myc. Strains YPH499 (SPT8+) and YMW220 (spt8Δ) were transformed with plasmids encoding the Gcn5∷13Myc (pVV231) or Spt7∷13Myc (pVV232) fusions and/or the TFIIS∷3HA fusion (pVV228). Protein extraction, immunopurification and detection were as for Figure 2A. No co-purification was observed when Gcn5∷13Myc and TFIIS∷3HA were expressed from their respective chromosomal locus (data not shown).