Fig. 4

FgESPs do not directly interact with DNMT1 or TET1 in buffalo DCs. a Western blot analysis for the Co-IP assay to determine the interaction between FgESPs and DNMT1 or TET1 protein in buffalo DCs. Lanes 1–3: protein-G beads-eluted ‘Ag-Ab-unknown interactive protein’ complex protein sample specifically bounded to anti-DNMT1 mAb (Lane 1), anti-TET1 mAb (Lane 2) or normal mouse IgG (Lane 3) as control Ab from FgESPs-treated DCs; Lanes 4–6: protein-G beads-eluted ‘Ag-Ab-unknown interactive protein’ complex protein sample specifically bounded to anti-DNMT1 mAb (Lane 4), anti-TET1 mAb (Lane 5) or normal mouse IgG (Lane 6) from untreated DCs. The laboratory-made FgESPs-specific pAb was used as a primary antibody for all lanes. The immunoblotting result showed that the band pattern among all protein samples had no significant difference. b Fluorescent subcellular localization for FgESPs in buffalo DCs observed under a fluorescence inverted microscope. DAPI (blue) stained the cell nucleus; lipophilic dye DiO (green) strongly bound to the cell membrane; and Cy3-pre-labelled FgESPs emitted orange fluorescence. The merged image demonstrated that FgESPs might not locate in the nucleus. Scale-bars: 20 μm