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. 2007 Mar 21;2(3):e316. doi: 10.1371/journal.pone.0000316

Figure 6. The TAF8-, TAF10- and SPT7L-containing complex has a size of about 300–350 kDa and may exist in vivo in crude nuclear extracts.

Figure 6

Immunupurified and eluted TAF10-containing complexes (TAF10 IP) (A), or HeLa nuclear extract (NE) (B) (50 µl) were injected on a Superose 6 size exclusion chromatography column using the SMART FPLC system (Pharmacia) and separated. Portions of the input (in; 5 µl) and every fraction from 7 to 38 (20 µl) were analysed by western blot (as indicated). Fraction numbers are shown under each lane and the elution profile of known molecular mass markers is indicated above the panels. To detect the non-phosphorylated form of SPT7L, 20 µl from each fraction was first phosphatase treated for 30 min at 37°C, then separated by SDS-PAGE and analyzed by western blot with the anti-SPT7L mAb (panel SPT7L-PPase). (C) Baculovirus expressed SPT7L in WCE (lane 2) and purified TFTC (lane 4) were treated with phosphatase (PPase) for 30 min at 37°C, separated on SDS-PAGE and analysed by western blot with the anti-SPT7L mAb (15) (upper panel), and an anti-TAF10 mAb (lower panel). The corresponding mock treated fractions are shown in lanes 1 and 3, respectively.