Fig. 7. Mammalian cohesin co-immunoprecipitates with ChlR1. HeLa cell lysates were incubated with protein G beads alone or in combination with pre-immune serum or ChlR1 2075 antibody as indicated at the top of the immunoblots. Specific interaction with components of the cohesin complex was detected by western blotting using antibodies to Smc1, Smc3, Scc1 or SA1/2 as indicated on the left. (B) To confirm specificity of the interaction, cells were transfected with 20 nM siRNAs as indicated at the top of the immunoblots. ChlR1 protein levels were efficiently reduced following transfection with specific siRNA but not with GFP control. Scc1 input is shown to be unaltered (middle panel). Depletion of ChlR1 protein levels results in a proportional decrease in the amount of co-immunoprecipitating Scc1 protein compared to controls (lower panel).