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Fig. 2. Effect of flusulfamide on the expression of immunophilin genes. (A) RT-qPCR analyses of 20 immunophilins (11 cyclophilins (PbCYP), seven FK506 binding proteins (FKBP), and two parvulin-like proteins (PAR) genes in P. brassicae resting spores treated with flusulfamide for 24 hr. Values represent means±S. E. Statistical significance was determined using Student’s t-test. Asterisks indicate significant differences between P. brassicae resting spores treated with water (control) and flusulfamide (1 ppm); *p<0.05 and **p<0.01. (B) Time–course gene expression analyses of three cyclophilin genes: PbCYP3 (open circles), PbCYP8 (filled squares), and PbCYP10 (open squares). Control (filled circles) corresponds to the reference gene (PbACT1) used for RT-qPCR normalization. Values represent means±S. E. Statistical significance was determined using Student’s t-test. Asterisks indicate significant differences between P. brassicae resting spores treated with water (control) and flusulfamide (1 ppm); *p<0.05 and **p<0.01. (C) Silver staining of SDS-PAGE (left) and immunoblot analysis (right) of proteins extracted from P. brassicae resting spores treated with flusulfamide for 24 hr: anti-PbCYP3 IgG was used as the primary antibody. C, flusulfamide-untreated; F, flusulfamide-treated.