Figure 1From: Rotavirus NSP4: Cell type-dependent transport kinetics to the exofacial plasma membrane and release from intact infected cellsNSP4 transport kinetics to the exofacial PM of HT29.F8 and MDCK cells vary. The cloned intestinal and canine kidney cells simultaneously were infected with RV SA11.4F (MOI = 2). At 2, 3, 4, 7, and 8 hpi, the cells were surface biotinylated at 4°C, lysed, and precipitated with streptavidin agarose. The cell lysates and precipitated surface proteins were separated by SDS-PAGE and analyzed by Western blot using NSP4150-175.peptide-specific antibodies (panels A and B). A cytofacial membrane (anti-caveolin-1) and intracellular (anti-NSP5) control are shown in panels C and D, respectively. Lanes were loaded as follows: Lane 1 = cell lysates recovered from the streptavidin pull-downs. Lane 2 = streptavidin precipitate of the surface biotinylated proteins. Lane 3 = cell lysates from a mock-biotinylated control. Lane 4 = streptavidin precipitate of the mock-biotinylated control, Lane 5 = cell lysates from an uninfected cell control.Back to article page