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Interaction analyses between OsMADS50 and OsMADS56. (a) Yeast two-hybrid assay. Two plasmids containing either AD or BD construct were introduced into yeast strain and transformants were grown on selective medium lacking Leu, Trp and His. OsMADS14-K bait and OsMADS1-KC prey were used as positive control (Lim et al. 2000). (b, c) Co-immunoprecipitation assay between OsMADS50 and OsMADS56 (b), and for OsMADS50 itself and OsMADS56 itself (c). Crude extracts were prepared from 7 DAG seedling protoplasts that had been transfected with constructs expressing HA-tagged protein or Myc-tagged protein, alone or with both. Transiently expressed proteins were immunoprecipitated with anti-Myc antibody, separated by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) then detected with peroxidase-conjugated high-affinity anti-HA antibody. Expected sizes of HA-tagged OsMADS50 and OsMADS56 were 34.05 and 34.18 kDa, respectively. Inclusion or exclusion of plasmid is shown by (+) or (−). DAG, days after germination.