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. 2019 Sep 1;33(17-18):1280–1292. doi: 10.1101/gad.325241.119

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© 2019 Yeom and Groisman; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 4. — Acetylated HspQ is neither a Lon substrate nor a Lon activator in vivo. (A) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQ-Flag qad (JY740), hspQK96R-Flag (JY892), hspQK96R-Flag qad (JY893), hspQK96Q-Flag (JY894), and hspQK96Q-Flag qad (JY895) Salmonella. (B) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQ-Flag qad (JY740), hspQ-Flag pat (JY898), and hspQ-Flag pat qad (JY899) Salmonella. (C) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQ-Flag qad (JY740), and hspQ-Flag qad pat (JY899) Salmonella with the plasmid vector (vector) or the qad-expressing plasmid (pqad) using 100 µM IPTG. (D) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQ-Flag qad (JY740), hspQ-Flag lon (JY703), and hspQ-Flag lon qad (JY745) Salmonella. (E) Western blot analysis of crude extracts from oat-Flag (JY655), oat-Flag lon (JY1009), oat-Flag hspQ (JY686), oat-Flag qad (JY774), oat-Flag lon hspQ (JY1011), hspQ qad (JY773), oat-Flag qad lon (JY1012), and lon hspQ qad (JY1013) Salmonella with a His-hha-expressing plasmid (pHis-hha). his-hha transcription from pHis-hha was induced with 100 µM IPTG. (F) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQ-Flag lon (JY703), hspQK96R-Flag (JY892), hspQK96R-Flag lon (JY2000), hspQK96Q-Flag (JY894), and hspQK96Q-Flag lon (JY2001) Salmonella. (G) Western blot analysis of crude extracts from hspQ-Flag (JY674), hspQK96R-Flag (JY892), and hspQK96Q-Flag (JY894) Salmonella with a His-hha-expressing plasmid (pHis-hha). his-hha transcription from pHis-hha was induced with 100 µM IPTG. Numbers below blots show quantification of the top band in the various strains. Words with italicized letters indicate gene names. For Western blot analysis, samples were analyzed with antibodies directed to acetylated lysine; the Flag, HA, or His epitopes; or the AtpB, OmpA, or GroEL proteins. Data are representative of three independent experiments, which gave similar results. See also Supplemental Figures S3 and S5.