Figure 5
Preferential sorting of NSP4 in polarized kidney and intestinal cells was distinct. MDCK and HT29.F8 cells were grown on permeable, membrane supports until differentiation as indicated by an elevated transepithelial resistance (TER, ≥ 500 ohms × cm2) and the presence of domes in cells grown on plastic. The cells were simultaneously infected with RV SA11.4F on apical membranes at MOI = 2 and the TER monitored to ensure the polarized phenotype was maintained 1 - 24 hpi. Shown in Panel A are the TER measurements of the infected MDCK and HT29.F8 cells taken at 1, 6, 15, 19, and 24 hpi. The x-axis indicates the hpi and the y-axis shows the corresponding % recovery of the pre-infected TER. Panel B. Apical or basolateral surfaces were biotinylated at 24 hpi with opposite surfaces treated with media. The cells were lysed, precipitated with streptavidin-agarose and subject to SDS-PAGE and Western Blot (Panel B). The M r markers are shown on the left. Lanes 1-4 are from the MDCK cells and lanes 5-8 from the HT29.F8 cells, with lanes 1 and 5 showing the apical surface, and lanes 2 and 6 the basolateral surfaces. Lanes 3-4 and 7-8 are the corresponding cell lysates.