Fig. 4

Small peptide can induce the degradation of fusion protein OVO::dsRED. a Svb/OVO N-terminal sequences from Drosophila (D_mel) and Bombyx mori (B_mor),K3, K8 and K28 respectively represent the third, the eighth and the twenty-eighth Lys residues at Svb/OVO N-terminus (b) Observation of red fluorescent cells. Tal and ovo::dsred, the cotransfected BmN cells with a full length of tal-like cDNA gene expression vector and DsRed fused with N- terminus 28 residues of BmOVO-1 expression vector. 5A1–4 + B and ovo::dsred, the cotransfected BmN cells with a sequence containing 5′-non coding sequence and it’s downstream 1A - 4A with B of tal-like cDNA gene expression vector and DsRed fused with N- terminus 28 residues of BmOVO-1 expression vector. A1–4 + B and ovo::dsred, the cotransfected BmN cells with a sequence containing A1-A4 with B of tal-like cDNA gene expression vector and DsRed fused with N- terminus 28 residues of BmOVO-1 expression vector. B and ovo::dsred, the cotransfected BmN cells with a sequence containing B and its downstream non coding sequence of tal-like cDNA gene expression vector and DsRed fused with N-terminus 28 residues of BmOVO-1 expression vector. Ovo::dsred, the transfected BmN cells with DsRed fused with N- terminus 28 residues of BmOVO-1 expression vector. NC means natural control representing BmN cells that do not transfect anything. (c) Western blotting of cells transfected with tal and ovo::dsred, B and ovo::dsred, 5A1–4 + B and ovo:: dsred, A1–4 + B and ovo::dsred, ovo::dsred. For detection of DsRed, the mouse antibody against RFP was used as a primary antibody, antibody goat anti-mouse HRP used as a secondary antibody. The tubulin was used as internal reference. 30 μg protein from the lysed cells was used for western blotting