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Figure 4 | Virology Journal

Figure 4

From: Mutational analysis of the rotavirus NSP4 enterotoxic domain that binds to caveolin-1

Figure 4

Western blot analyses of caveolin. Yeast lysates expressing Rev2; Rev2 116, 124,127A; Rev2 116A; Rev1,2 127A; Rev1,2 124,127A, and NSP4HydroMut were incubated with CNBr-activated sepharose 4B beads bound by either the N-terminal (cav-119-40) (lanes 1-7) or C-terminal (cav-1161-178) (lanes 8-14) peptides. Left Panel: Yeast lysates were reacted with bound cav-1 residues 19-40 and probed with rabbit anti-NSP4150-175. Lanes 1, 4, and 5 depict the reactivity of Rev2; Rev1,2 127A; and Rev1,2 124,127A, respectively. Lanes 2, 3, 6, and 7 show a lack of reactivity with Rev2 116, 124,127A; Rev2 116A; NSP4HydroMut; and InVSc1 respectively. Right Panel: The same binding pattern was observed when the yeast lysates were incubated with cav-1161-178 (lanes 8-14). RV-infected MDCK cell lysates (lane 15) demonstrate the specificity of the rabbit anti-NSP4150-175. Non-specific binding to the beads alone was not observed (data not shown). Lanes 4, 5, 11, and 12 show multimeric forms of NSP4.

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