Specific binding of rTsAP with IEC and cellular localization. A IFT analysis of specific binding of rTsAP with IEC. The IEC or C2C12 cells were pre-incubated with rTsAP or PBS. After blocking and washes, the IEC and C2C12 were incubated with anti-rTsAP serum, infection serum or pre-immune serum, followed by the incubation of FITC-conjugated anti-mouse IgG. Cell nuclei were re-dyed red by propidium iodide (PI). Immunostaining was observed on the surface of IEC probed by anti-rTsAP serum and infection serum, but not by pre-immune serum. Scale-bars: 25 μm. B Cellular localization of rTsAP in IEC by confocal microscopy. The IEC were first pre-incubated with rTsAP, subsequently probed using anti-rTsAP serum, infection serum or pre-immune serum, finally stained using FITC-conjugated anti-mouse IgG. PI was utilized to dye cell nuclei red. Immunostaining was principally located within IEC cytoplasm. Scale-bars: 5 μm.