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. 2016 May 13;11(5):e0155539. doi: 10.1371/journal.pone.0155539

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© 2016 Kim et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 4 — In vivo repressor activities of PerR_BL, Fur_BL, and Zur_BL (A) Repressor activities of PerR_BS and PerR_BL for P_mrgA-lacZ reporter fusion. B. subtilis cells expressing no PerR orthologue (LB1532), PerR_BS-FLAG (HB9738), or PerR_BL-FLAG (LB1023) were treated without or with 100 μM H₂O₂ for 30 min, and β-galactosidase activities were measured using P_mrgA-lacZ reporter fusion. (B) Repressor activities of Fur_BS and Fur_BL for P_feuA-lacZ reporter fusion. B. subtilis cells expressing no Fur orthologue (LB1040), Fur_BS-FLAG (LB1041), or Fur_BL-FLAG (LB1042) were treated without or with 100 μM H₂O₂ for 30 min, and β-galactosidase activities were measured using P_feuA-lacZ reporter fusion. (C) Repressor activities of Zur_BS and Zur_BL for P_yciC-lacZ reporter fusion. B. subtilis cells expressing no Zur orthologue (LB1034), Zur_BS-FLAG (LB1035), or Zur_BL-FLAG (LB1036) were treated without or with 100 μM H₂O₂ for 30 min, and β-galactosidase activities were measured using P_yciC-lacZ reporter fusion. (D-F) Western blot analyses of FLAG-fused PerR orthologues (D), Fur orthologues (E), and Zur orthologues (F). The FLAG-fused proteins were probed by anti-FLAG antibody. (G) Fe-dependent repressor activities of Fur_BS and Fur_BL for P_feuA-lacZ reporter fusion. B. subtilis cells expressing no Fur orthologue (LB1040), Fur_BS-FLAG (LB1041), or Fur_BL-FLAG (LB1042) were grown in MLMM supplemented with or without 10 μM FeSO₄, and β-galactosidase activities were measured using P_feuA-lacZ reporter fusion. (H) Zn-dependent repressor activities of Zur_BS and Zur_BL for P_yciC-lacZ reporter fusion. B. subtilis cells expressing no Zur orthologue (LB1034), Zur_BS-FLAG (LB1035), or Zur_BL-FLAG (LB1036) were grown in MLMM supplemented with or without 10 μM ZnCl₂, and β-galactosidase activities were measured using P_yciC-lacZ reporter fusion.