As09532a

Protein samples were separated by SDS-PAGE gel and transferred to PVDF membranes.
Antibodies against FCA 39 , SE (Agrisera, AS09532A), HYL1 40 (link) , ELF3 41 (link) , H3 (Abcam, ab1791), GFP (Roche, 11814460001), Tubulin (Sigma, T5168) were used as primary antibodies. After the primary antibody incubation, horseradish peroxidase (HRP)conjugated secondary antibodies (GE Healthcare) were used for protein detection by chemiluminescence (Thermo Scientific, 34095).

Western blots were performed as described 65 . Proteins from 14-day-old seedlings were extracted, resolved in 12% (v/v) SDS-polyacrylamide gels, and transferred to Hybond C-Extra membranes (Amersham Biosciences). The membranes were blocked with 5% (w/v) non-fat milk in Tris buffered saline Tween (TBST) buffer and then probed with speci c antibodies. Antibodies used included anti-GAPDH (Santa Cruz Biotechnology, sc-365062, dilution, 1:1000), anti-SUL (dilution, 1:1000) 36 , anti-HYL1 (Agrisera, AS06136, dilution, 1:2000), anti-AGO1 (Agrisera, AS09527, dilution, 1:2000), anti-SE (Agrisera, AS09532A, dilution, 1:1000), and anti-GFP (Abcam, ab290, dilution, 1:3000). After three washes, the membranes were probed with horseradish peroxidase-conjugated, goat-anti-rabbit IgG (Bio-Rad, cat.172-1019) (dilution, 1:2000) or goat-anti-mouse IgG (Bio-Rad, cat.170-6516, dilution, 1:2000). The protein signals were detected with the Amersham TM ECL TM Prime Western Blotting Detection Reagent (GE healthcare, RPN2232) and visualized with the Chemiluminescence imaging system (Clinx Science Instruments Co. Ltd, China).