Fig. 5.

Identification of the SBP1 protein in apple. (A) Y2H analysis between MdSBP1 and different haplotype of S-RNases. S ₁-, S ₂-, S ₃-, S ₅-, S ₇-, and S ₉-RNase represent the full length, while S-RNase-C2HVC3 and S-RNaseC4C5 represent the hypervariable and conserved region, respectively. The detail of this is in Supplementary Fig S7 at JXB online. (B) Y2H analysis between MdSBP1 and MdCUL1, S2-MdSFBB1–4, and S2-MdSFBL1–6. (C) The interaction analysis between MdSBP1 and MdSSK1. The AD fusions and BD fusions were co-transformed into yeast strain AH109. Transformants were grown on selective medium SD/-Ade-His-Leu-Trp for 7 d at 30 ° and then were stained by X-α-gal. The p53 and SV40 were used as positive controls, the pGBKT7 and pGADT7 as negative controls. (D) Pull-down analysis of MdSBP1 and MdCUL1. Purified MBP-MdCUL1 was used as bait against purified GST-MdSBP1. Bound proteins were examined with anti-GST antibody. GST and MBP were used as negative controls.