1. Volume 14, Issue 5, Pages 685-691 (June 2004)
A Set of Consensus Mammalian Mediator Subunits Identified by Multidimensional Protein Identification Technology 
Shigeo Sato, Chieri Tomomori-Sato, Tari J Parmely, Laurence Florens, Boris Zybailov, Selene K Swanson, Charles A.S Banks, Jingji Jin, Yong Cai, Michael P Washburn, Joan Weliky Conaway, Ronald C Conaway 
Molecular Cell 
Volume 14, Issue 5, Pages (June 2004)
DOI: /j.molcel

2. Figure 1 Consensus Mammalian Mediator Subunits Identified by MudPIT
(A) Mammalian Mediator subunits identified in different Mediator preparations. Mediator subunits detected in the TRAP/SMCC, ARC, DRIP, CRSP, PC2, mouse Mediator, and rat Mediator are indicated in red. Proteins not detected are indicated in green. (B) Mammalian Mediator subunits identified by MudPIT analyses of Mediator preparations from FLAG-Nut2 (MED10), FLAG-Med25 (MED9), FLAG-Intersex (MED29), FLAG-LCMR1 (MED19), or FLAG-MED28-expressing HeLa cell lines. A portion of the MudPIT data set for the FLAG-Med25 Mediator preparation was described elsewhere (Tomomori-Sato et al., 2003). (C) Analyses of Mediator preparations following phosphocellulose chromatography. P0.5 and P1.0 correspond to fractions eluted from phosphocellulose from 0.3 to 0.5 M KCl and from 0.5 to 1.0 M KCl, respectively. In (B) and (C), Mediator subunits detected by MudPIT are indicated in red with the percent amino acid sequence coverage; proteins not detected are indicated in green. “*,” a single potential TRAP230 peptide was identified by MudPIT in the P1.0 fraction but could not be unambiguously assigned to TRAP230.
Molecular Cell  , DOI: ( /j.molcel )

3. Figure 2
SDS Polyacrylamide Gel of Mediator Preparations Analyzed by MudPIT
Aliquots of anti-FLAG agarose eluates prepared from equivalent numbers of parental HeLa cells and HeLa cells stably expressing FLAG-Nut2 (MED10) [f:Nut2 (MED10)], FLAG-Intersex (MED29) [f:Intersex (MED29)], FLAG-Med25 (MED9) [f:Med25 (MED9)], FLAG-LCMR1 (MED19) [f:LCMR1 (MED19)], FLAG-MED28 (f:MED28), or FLAG-CRSP70 (MED26) [f:CRSP70 (MED26)] were subjected to SDS polyacrylamide gel electrophoresis in a 4%–20% gradient gel visualized by silver staining. The positions and relative molecular masses in kDa of protein size standards for lanes 1–6 are indicated on the left. Lane 9 shows size markers for lanes 7 and 8.
Molecular Cell  , DOI: ( /j.molcel )

4. Figure 3 Pol II Subunits Identified in Mediator Preparations by MudPIT
Pol II subunits detected by MudPIT are indicated in red with the percent amino acid sequence coverage; polymerase subunits not detected are indicated in green.
Molecular Cell  , DOI: ( /j.molcel )

5. Figure 4
TRAP230 and Cdk8 Are Underrepresented in Mediator Purified through CRSP70
Aliquots of anti-FLAG agarose eluates from HeLa cells expressing FLAG-Nut2 (MED10) [f:Nut2 (MED10)] or FLAG-CRSP70 (MED26) [f:CRSP70 (MED26)] were analyzed by immunoblotting using antibodies against MED8 (Brower et al., 2002), TRAP230 (MED12) (S-19, Santa Cruz Biotechnology), CDK8 (D-9, Santa Cruz Biotechnology), or Rpb1 (N-20, Santa Cruz Biotechnology).
Molecular Cell  , DOI: ( /j.molcel )