Fig.1 Identification of RBP2 in crude cell extracts by Western blotting with antibody 18E8.
Samples: 1. HeLa control siRNA 2. HeLa RBP2 siRNA 3. MCF7 4. U2OS 5. NIH3T3
6. J1 (mouse ES)
Fig.2 Immunofluorescence staining of HeLa cell with anti-RBP” antibody.
1. HeLa cells were fixed with 4% paraformaldehyde overnight, permealized with 0.25% Triton X-100 in PBS for 10 min.
2. Incubate cells with 1.5% BSA in PBS for 30 min to block non-specific binding of the antibodies. Incubate the cells with 1/2,000 diluted anti-RBP2 antibody (18E8) in 1% BSA in PBS at 4°C overnight.
3. Incubate cells with a secondary antibody, goat anti-mouse IgG conjugated with Alex 488, at 1/1,000 dilution in 1% BSA for 1 hr at room temperature.
4. Nucleus (DNA) was stained with DAPI