Figure 2.

RAD21 Alteration Effects on the Cohesin Complex

(A) RAD21 p.Pro376Arg increases binding affinity to mitotic STAG proteins. Immunoblotting of similarly expressed hybrid proteins is indicated below. Significant p values (<0.05) from two-tailed t tests are indicated (^∗). Open (B), partially open (C) and closed (D) sister-chromatid phenotypes are demonstrated. (E) Increased prevalence of closed sister chromatids in LCLs from the boy with the p.Pro376Arg mutation and in normal control LCLs (C1, C2). The RAD21-deleted LCLs (del) show no difference from controls. The number of metaphases analyzed for each cell line is indicated in white at the base. Chi-square calculations of significance in difference of all cell phenotypes of RAD21 mutant cell lines compared with the averaged controls are indicated above the columns. (F) Flow cytometry smoothened histogram plots of BrdU-positive cells versus DNA content showing the proportions of G1, S, and G2/M cells 0, 6, and 21 hr after BrdU labeling. The proportion of cells in each cell cycle phase is expressed as the percentage of BrdU-positive cells. Two-sided Fisher's exact test is shown. Error bars indicate standard deviations. del = RAD21 deletion, P376R = p.Pro376Arg, C585R = p.Cys585Arg mutations, and CON = control cells.