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The d2 domain is required for Pet internalization and epithelial cell damage. HEp-2 cells were treated with Pet or its mutants (PetΔd1, PetΔd2, or PetΔPDWET) or Pet d1 at 20 μg/ml for 3.5 h. The cells were washed, fixed, permeabilized, and stained with rhodaminated phalloidin for F actin (red), TO-PRO 3 for DNA (blue), and anti-Pet antibodies, followed by a secondary anti-rabbit IgG antibody labeled with fluorescein (green). The slides were observed using confocal microscopy.
