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<t>(A–D)</t> <t>ChIP-seq</t> profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.
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<t>(A–D)</t> <t>ChIP-seq</t> profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.
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<t>(A–D)</t> <t>ChIP-seq</t> profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.
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<t>(A–D)</t> <t>ChIP-seq</t> profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.
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Image Search Results


(A–D) ChIP-seq profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.

Journal: bioRxiv

Article Title: Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica

doi: 10.64898/2026.02.23.707304

Figure Lengend Snippet: (A–D) ChIP-seq profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.

Article Snippet: Immunoprecipitated DNA was subjected to ChIP-seq library preparation using Novogene’s standard workflow, including end repair, A-tailing, Illumina adapter ligation, size selection, and PCR amplification, and sequenced by Novogene (Munich, Germany) on an Illumina NovaSeq X Plus platform with paired-end 150 bp reads.

Techniques: ChIP-sequencing, Binding Assay, Control, Membrane, Permeability, Expressing, Gene Expression, Derivative Assay, RNA Sequencing, Activity Assay, Fluorescence, Mutagenesis