Supplemental Material

This article contains the following supporting material:

• Figure 1 - Recombinant human BAF phosphorylated in S-phase Xenopus extracts resolves into four acidic-shifted, phosphorylated species. Isolated, untagged (thrombin-cleaved) human BAF dimers were incubated in S-phase Xenopus extracts in the presence of ³²PγATP (as in Fig. 2) and resolved on pH 4-7 linear IPG strips in the first dimension and SDS-PAGE in the second dimension. The recombinant, thrombin-cleaved BAF monomer is predicted to migrate at pI 6.13 (indicated by dotted line) and has a predicted mass of 10.4 kDa. The autoradiography image of the gel is shown and the acidic-shifted BAF spots are indicated by arrows. We attribute the two ‘extra’ spots seen here (compare to Fig. 1B, HeLa) to the higher resolution of the IEF strips used and more sensitive detection of ³²P (rather than immunoblotting). The acidic-shifted spots all correspond to phosphorylated BAF, however they can originate from any combination of phosphorylation/acetylation events. We speculate that the major spot corresponds to BAF phosphorylated on Ser-4.

• Table 1 - Primers and vectors used for cloning. The different BAF constructs listed above were made either by amplifying the BAF cDNA by PCR and cloning it into appropriate vectors using restriction enzymes BamHI and XhoI (constructs 2, 4 and 7), by cutting out suitable BAF cDNA sequences from other vectors using BamHI and XhoI and ligating into target vectors (‘copy and paste’; constructs 5 and 6), or by site-directed mutagenesis (constructs 1, 3, 8-11).