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. 2020 Jun 16;11(3):e00363-20. doi: 10.1128/mBio.00363-20

FIG 6.

FIG 6

Hfq, sRNA 0161, and sRNA 179 inhibit ExsA synthesis. (A) WT PA103 and an exsA mutant carrying a rhamnose-inducible copy of exsA integrated at the Tn7 site were cultured under noninducing (- EGTA) or inducing (+ EGTA) conditions for T3SS gene expression. Rhamnose (0.005%) was included in the growth medium to induce exsA expression from the Tn7 integrant, and ExsA activity and protein levels were measured using the PexsD-lacZ reporter and ExsA immunoblots, respectively. (B) The indicated strains carrying the rhamnose-inducible copy of exsA were transformed with either the vector control, Hfq expression vector, sRNA 1061, or sRNA179 expression plasmids, cultured under T3SS inducing conditions, and assayed for PexsD-lacZ reporter activity and ExsA protein levels. (C) Comparison of exsA mRNA and ExsA protein levels in samples harvested from an exsA mutant carrying the rhamnose-inducible copy of exsA and either a vector control, pHfq, p0161, or p179. RNA samples were quantified for exsA using qRT-PCR and normalized to the rimM housekeeping gene. The reported values are relative to the exsA strain carrying the vector control.