Figure 5.

Protein levels in cross-linked whole-cell extract or chromatin samples over time. A, Western blots of WCE samples for all factors except Gal4, which is a chromatin extract. Antibodies used are listed in supplemental Table S4, and molecular mass is denoted to the right in kDa. Samples were cross-linked with 5% formaldehyde for 0–15 min and quenched with excess glycine. B, quantification of WCE Western blot bands shown in A for TBP. Each sample was normalized to the zero time point as a percentage. Two replicates were averaged for the plot, and error bars represent standard deviation. C, same as B, except for Ace1. D, same as B, except for transcription factors Reb1, Cat8, Abf1, Sir2, and Sko1. E, same as B, except for preinitiation complex components TFIIB, Tfa1 (TFIIE), Tfg1 (TFIIE), and Rpb1 (RNA polymerase II). Independently performed Western blottings using chromatin rather than WCE samples showed the same trends. F, same as B, except for Gal4 chromatin extract. Gal4 was not abundant enough to be detected in WCEs.