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[Preprint]. 2023 May 8:2023.05.07.539761. [Version 1] doi: 10.1101/2023.05.07.539761

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Figure 7. — Evidence for a monomeric state of Alx and MntP in vivo. (A) Immunoblots probed with anti-HA antibody after electrophoretic mobility of samples containing crude membrane preparations. The crude membranes were harvested for strains RAS31, RAS32, and RAS130 expressing Alx^HA, MntP^HA, and MscL^HA, respectively, and treated with a crosslinker DSS. (B) β-galactosidase activity (Miller units) as a reporter of alx translation (P alx-PRE-alx’-lacZ, pRA54) was measured in mid-log phase grown cultures of alx⁺ strain (MC4100) bearing a vector (pHYD5001) or a derivative of pHYDSOOl expressing Alx^HA (pRA27), Alx^HA_D92N (pRA62), AIx^HA_E213Q (pRA63), or Alx^HA_D216N (pRA64) from a Ptrc promoter. The cultures were grown in LBK media with pH 6.8 and 8.4, supplemented with appropriate concentration of ampicillin and IPTG. The expected outcome of the experiment is summarized in the table. The error shown is standard deviation of three repeats of the experiment.