Supplementary Figure 1: Mutational analysis of the SA2-Scc1 interaction in vitro and in human cells.
From: Structure of cohesin subcomplex pinpoints direct shugoshin-Wapl antagonism in centromeric cohesion
(a) Autoradiograph (top) and Coomassie stained gel (bottom) of 35S-labeled Myc-SA2 proteins (input) and the same proteins bound to GST or GST-Scc1 beads. WT, wild type. (b) Quantification of the in vitro binding assays in a. The binding intensities were normalized to the amount of input for each SA2 protein. Error bars, s.d. (n = 3, independent experiments). The Y331A, Y479A, K1009E L1010A mutants were tested twice. Only the means were shown for these samples. (c) Autoradiograph (top) and Coomassie stained gel (bottom) of 35S-labeled Scc1-Myc proteins (input) and the same proteins bound to GST or GST-SA2 beads. WT, wild type. (d) Quantification of the in vitro binding assays in c. The binding intensities were normalized to the amount of input for each Scc1 protein. Error bars, s.d. (n = 3, independent experiments). (e) Anti-Myc or anti-GFP immunoblots of lysates and anti-Myc IP of HeLa cells co-transfected with plasmids encoding GFP-SA2 and the indicated Myc-Scc1 proteins. WT, wild type.
