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. 2019 Sep 10;10(1):783–792. doi: 10.1080/21505594.2019.1661721

Figure 3.

Figure 3.

Ler mediates activation of LEE genes by OvrB in O157. (a-e) EMSA of the specific binding of OvrB to PLEE1 (a) rpoS (negative control) (b), PLEE2/3 (c), PLEE4 (d), and PLEE5 (e). PCR products were added to the reaction mixtures at 40 ng each. OvrB protein was added to the reaction buffer in lanes 2–7 at 0.1, 0.2, 0.4, 0.8, 1, and 2 μM, respectively. No protein was added in lane 1. (f) Fold enrichment of the LEE1, LEE2/3, LEE4, and LEE5 promoters in OvrB ChIP samples, as measured by qPCR. rpoS served as negative control. (g) Adherence of O157 WT, ΔLer mutant, and ΔLerΔovrB double mutant to HeLa cells. (h) qRT-PCR analysis of changes in LEE gene expression in O157, ΔLer mutant, and ΔLerΔovrB double mutant. Data represent mean ± SD (n = 3). *P ≤ 0.05; **P ≤ 0.01, ***P ≤ 0.001 (Student’s t-test).