Validation of proteomics results by Western blot analysis.

To validate proteomics results, selected differentially represented immune system proteins were produced in E. coli and used to generate antibodies in rabbits for Western blot analysis of individual wild boar mandibular lymph node protein samples (young TB-, N = 5; young TB+, N = 9; adult TB-, N = 4; adult TB+, N = 5; adult TB++, N = 5). The intensity of protein bands corresponding to test and control RPS14 proteins was determined by densitometric analysis. The intensity of test protein bands was normalized against the intensity of the control band, represented as average + S.D. and compared between groups in adult or young wild boars by a multivariate comparison between the groups using the one-way ANOVA test followed by one-tailed Student’s t-test with Bonferroni correction for samples with unequal variance (p = 0.05). (A) Normalized S100A9 protein levels. (B) Normalized LTF protein levels. (C) Normalized PGLYRP1 protein levels. (D) Comparative analysis of proteomics and Western blot results for differentially represented immune system proteins S100A9, LTF and PGLYRP1.