FIG. 5.

Interaction of hSNM1A and hPIAS1. (A)Yeast two-hybrid assay. Yeast cells were cotransformed with the bait plasmid containing either hSNM1AΔ1-554 or its derivatives (with the D838N or H994A mutation) and the prey plasmid containing hPIAS1. Empty bait plasmid was used for control. β-Galactosidase activity was determined by a liquid culture assay and calculated in Miller units. (B and C) 293T cells were cotransfected with the indicated expression plasmids. Ni-NTA resin pull-down analysis was carried out as described in Materials and Methods. Samples were analyzed by Western blotting with anti-GFP or anti-His antibodies.