The pull-down assays were performed as previously described (39): 3 μg of fusion protein was bound to beads. These beads were incubated overnight with soluble protein at 4°C and then washed. SDS–polyacrylamide gel electrophoresis (PAGE) was used to resolve the proteins, which were detected by immunoblotting with anti-GST (A00866-100, lot: 13D000626; GenScript) or anti-His (M30111M, lot: 273884; Abmart).

The Co-IP assays were also conducted as previously described (39, 40). The proteins of interest—SSU72, FCA, and CLF—were tagged with FLAG or HA, and the fusion constructs were cloned into pUC19. The constructs were transformed into protoplasts from Arabidopsis; the protoplasts were then incubated overnight at 22°C. Protein extracts from lysed protoplasts were immunoprecipitated with anti-FLAG (H6908, lot: SLBQ7119V; Sigma-Aldrich) or anti-HA (H9658, lot: 095M4778V; Sigma-Aldrich). After washing the beads, the immunoprecipitated proteins were resolved by SDS-PAGE and detected by immunoblotting with anti-HA or anti-FLAG.

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