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. Author manuscript; available in PMC: 2019 May 28.
Published in final edited form as: Cell Rep. 2019 May 21;27(8):2468–2479.e3. doi: 10.1016/j.celrep.2018.12.063

Figure 4. Induction of HtrA-LppZ Decreases Cellular Ami3 Levels.

Figure 4.

(A) HtrA and LppZ expression decrease Ami3 levelsto varying degrees. Strains in which the only copy of htrA or lppZ was under an aTc-inducible promoter were constructed and grown with or without 100 ng/mL aTc for 12 hr. HtrA(S354A) is an allele of htrA presumed to exhibit reduced catalytic activity in HtrA homologs due to a mutation in its catalytic serine (see Figure S3). Whole-cell lysate was analyzed using western blotting using anti-Strep and anti-RpoB as a loading control.

(B) Higher expression of HtrA(S354A) further decreases Ami3. A second copy of aTc-inducible htrA(S354A) was transformed into the mutant strain described in (A), and all strains were grown with or without 100 ng/mL aTc for 8 hr. Whole-cell lysate was analyzed using western blotting using anti-Strep and anti-HtrA and anti-RpoB as a loading control. Western blot images were cropped but display all relevant lanes and reactive bands.