Fig. 2

Physical interaction between OsUev1s and OsUbc13. a Physical interaction between OsUev1s and OsUbc13 by a yeast two-hybrid assay. PJ69-4A transformants carrying one Gal4_AD (from pGAD424 derivative) and one Gal4_BD (from pGBT9 derivative) were replicated onto various plates as indicated and incubated at 30 °C for 3 (or 5, as indicated) days before being photographed. b Protein interactions between OsUev1s and OsUbc13 by an affinity pull-down assay. E. coli BL21 cells transformed with GST-OsUev1D (Lane 2), GST (Lane 8) and His₆-OsUbc13 (Lane 9) alone served as controls. pGEX6 (GST) co-transformed with His₆-OsUbc13 (Lane 7) also served as a negative control. Lane 1 contains prestained molecular weight markers (SM0661, Fermentas). Lanes 2–8 contain samples from the GST affinity pull-down products while Lane 9 contains purified His₆-OsUbc13. The SDS-PAGE gel was stained with Coomassie blue