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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Mol Microbiol. 2014 Feb 8;91(5):976–995. doi: 10.1111/mmi.12509

Fig. 6. EmrA1 mutant fails to secrete antioxidant enzymes SodB and KatG.

Fig. 6

The cultures of F. tularensis (Ft) LVS, the emrA1 mutant or the transcomplemented strain (emrA1+ pemrA1) were grown in MH- (A) or BHI-broth (B). The culture filtrates or the lysates of the bacterial pellet were analyzed at the indicated times by western blot analysis using anti-SodB and anti-KatG antibodies.

(C) Aliquots of the bacterial strain grown in MHB or BHI were collected at the indicated time points, diluted 10-fold and plated on MH-chocolate agar plates to enumerate bacterial numbers (n=3).

(D) The western blots of the MH-B or BHI-grown culture filtrates probed with anti-SodB antibodies were stripped and re-probed with antibodies against 50S ribosomal protein Rp1L at the indicated times (top panel). Agarose gel electrophoresis of the genomic DNA isolated from culture filtrates from the BHI-grown bacteria at indicated time points (lower panel).

(E) Determination of catalase activity of secreted KatG by amplex red assay. The amount of H2O2 consumed by culture filtrates of Ft LVS, the emrA1 mutant and the transcomplemented strain collected at the indicated times. The data are representative of at least 3–5 independent experiments conducted with identical results. The data were analyzed by ANOVA with a Tukey-Kramer post-test, and a cut-off p value of 0.05 or less was considered significant. Comparisons are shown with Ft LVS. ***, p<0.001.