Effect of PPARγ agonists and ATRA on ECwt infectivity in small intestinal villi. (a) Immunocytochemistry analysis using antibodies against viral structural proteins (SP) and nonstructural proteins (NSP4 and NSP5) present in villi from in vivo ECwt-infected mice (n = 3 per experiment) that had been treated or not with PGZ. (b) Immunocytochemistry analysis as in (a), except that villi isolated from mice (n = 3 per experiment) were infected in vitro with ECwt and treated or not with PGZ. (c) Capture ELISA analysis of lysates from in vitro ECwt-infected villi that had been treated or not with PGZ, RGZ, TZD, ALA, DHA, or ATRA. Viral antigen was detected with anti-SP antibodies (Ab). (d) Infectious titers of lysates from in vitro ECwt-infected villi that had been treated with PGZ. Viral titers were determined by immunochemistry assay using anti-SP antibodies. (e) Infectious titers of lysates from in vitro ECwt-infected villi that had previously been inoculated with the villus lysates analyzed in (d) and treated again with PGZ. (f) Infectious titers of lysates from in vitro ECwt-infected villi that had previously been inoculated with lysates from villi treated with RGZ, TZD, ALA, DHA, or ATRA and then treated with these same PPARγ agonists. (g) Cell viability of villus cells determined with the trypan blue exclusion test. H2O2 was used as a control. Data are presented as mean ± SD from three independent experiments performed in duplicate.