Figure legends
Figure 1Schedules of therapeutic (A), prophylactic (B), vaccination and grouping (C) for For t 2 DNA with skin patches. Time points of sensitization with rFor t 2/alum, videotaping, and sacrificing are indicated.
Figure 2 Scratching bouts, changes in serum For t 2-specific IgE and IgG1/IgG2a ratio, and mRNA expression levels of IL-13, interferon γ (INF-γ), IL-10 and Foxp3 in For t 2-stimulated splenocytes from (A) therapeutic and (B) prophylactic groups by real-time PCR. The statistical significance of differences between groups was assessed by the Bonferroni multiple range test. * denotes p<0.05, ** denotes p<0.01, ns denotes not statistically significant.
Figure S1(A) Dose finding schedule of For t 2 DNA patch vaccine on rFor t 2-sensitized mice in the pilot study. (B) Counts of scratching bouts from each group of mice before (day 0) and after (day 61) challenged by intradermal injection of rFor t 2. (C) Total IgE and For t 2-specific IgG2a antibodies in the sera at week 9 determined by ELISA. * denotesp <0.05, **denotesp <0.01, *** denotesp <0.001, ns denoted not statistically significant by the Bonferroni multiple range test.
Figure S2 The representative skin images show before (A) and after (B) patch application. The For t 2 DNA vaccine was delivered onto the pre-shaved abdominal skin through a Fin chamber for 1 hour.
Figure S3 H&E and immunohistochemotry (IHC) staining using rabbit anti-For t 2 polyclonal antibody of skin and spleen sections under 400x light microscope. For t 2 protein expression of skin and spleen were examed 24 and 48 hours after the last treatment of epicutaneous vaccination.
Figure S4 Effects of For t 2 vaccine on histopathology of challenged skins from (A) therapeutic or (B) prophylactic groups by H&E staining. Figures showed the representative abdominal skin sections obtained 48 hours after intradermal challenge under 100 x and 400 x light microscope. Moreover, the infiltrating inflammatory cells were quantified under 400-fold view from each group. The statistical significance of differences between groups was assessed by the Bonferroni multiple range test. ** denoted p<0.01, ns denotes not statistically significant.