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( A ) Representative microscopy images of wild-type (WT) (top) and dnaC2 (bottom) cells <t>expressing</t> <t>HU-mCherry</t> growing under microscope observation in M9glyCAAT. DNA replication in dnaC2 cells was blocked by growing cells at 37°C for 145 min. Arrows indicate dnaC2 cells with one (1N) or two (2N) nucleoids. Scale bars: 1 µm. ( B ) Graph showing the percentage of dnaC2 cells (CJW7374) with 1, 2, or >2 nucleoids after growth at 37°C. Shown are aggregated data from <t>three</t> biological replicates.
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( A ) Representative microscopy images of wild-type (WT) (top) and dnaC2 (bottom) cells <t>expressing</t> <t>HU-mCherry</t> growing under microscope observation in M9glyCAAT. DNA replication in dnaC2 cells was blocked by growing cells at 37°C for 145 min. Arrows indicate dnaC2 cells with one (1N) or two (2N) nucleoids. Scale bars: 1 µm. ( B ) Graph showing the percentage of dnaC2 cells (CJW7374) with 1, 2, or >2 nucleoids after growth at 37°C. Shown are aggregated data from <t>three</t> biological replicates.
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Image Search Results


( A ) Representative microscopy images of wild-type (WT) (top) and dnaC2 (bottom) cells expressing HU-mCherry growing under microscope observation in M9glyCAAT. DNA replication in dnaC2 cells was blocked by growing cells at 37°C for 145 min. Arrows indicate dnaC2 cells with one (1N) or two (2N) nucleoids. Scale bars: 1 µm. ( B ) Graph showing the percentage of dnaC2 cells (CJW7374) with 1, 2, or >2 nucleoids after growth at 37°C. Shown are aggregated data from three biological replicates.

Journal: eLife

Article Title: Genome concentration limits cell growth and modulates proteome composition in Escherichia coli

doi: 10.7554/eLife.97465

Figure Lengend Snippet: ( A ) Representative microscopy images of wild-type (WT) (top) and dnaC2 (bottom) cells expressing HU-mCherry growing under microscope observation in M9glyCAAT. DNA replication in dnaC2 cells was blocked by growing cells at 37°C for 145 min. Arrows indicate dnaC2 cells with one (1N) or two (2N) nucleoids. Scale bars: 1 µm. ( B ) Graph showing the percentage of dnaC2 cells (CJW7374) with 1, 2, or >2 nucleoids after growth at 37°C. Shown are aggregated data from three biological replicates.

Article Snippet: Fluorescence emission was collected during a 100 or 200 ms exposure time provided by a Spectra III Light Engine LED excitation source (Lumencor): mCherry—594 nm excitation, DAPI/FITC/TxRed filter cube (polychroic FF-409/493/596-Di02, triple-pass emitter FF-1-432/523/702-25), GFP and SYTO RNASelect—488 nm excitation, DAPI/FITC/TxRed polychroic filter cube, and an ET525/50M emission filter; YFP—514 nm excitation, CFP/YFP/mCherry filter cube (polychroic FF-459/526/596-Di01, triple-pass emitter FF-1-475/543/702-25), and a FF02-525/40-25 emission filter.

Techniques: Microscopy, Expressing

( A ) Images of representative cells from a mixed population of 1N (CRISPR interference [CRISPRi] oriC ) and multi-N (CRISPRi ftsZ ) cells. Strains CJW7457 and CJW7576 carrying HU-mCherry were used for the SYTO RNASelect staining experiment, whereas DAPI-stained strains SJ_XTL676 and SJ_XTL229 were used for the EUB338 ribosomal RNA (rRNA) fluorescence in situ hybridization (FISH) experiment. ( B ) Concentration distribution of SYTO RNASelect (3077 cells for each population from five biological replicates) and EUB338 (1254 cells for each population from three biological replicates) in 1N and multi-N cells. ( C ) The average 1N/multi-N SYTO RNASelect and EUB338 concentration ratio (gray bar) calculated from five and three biological replicates (white circles), respectively. ( D ) RNASelect and EUB338 concentration ratios as functions of cell area (mean ± SD from five and three biological replicates, respectively). Single exponential decay functions were fitted to the average ratios ( R 2 >97%) for each indicated reporter. All concentration comparisons or ratio calculations were performed for equal numbers of 1N and multi-N cells and overlapping cell area distributions (see Materials and methods and ).

Journal: eLife

Article Title: Genome concentration limits cell growth and modulates proteome composition in Escherichia coli

doi: 10.7554/eLife.97465

Figure Lengend Snippet: ( A ) Images of representative cells from a mixed population of 1N (CRISPR interference [CRISPRi] oriC ) and multi-N (CRISPRi ftsZ ) cells. Strains CJW7457 and CJW7576 carrying HU-mCherry were used for the SYTO RNASelect staining experiment, whereas DAPI-stained strains SJ_XTL676 and SJ_XTL229 were used for the EUB338 ribosomal RNA (rRNA) fluorescence in situ hybridization (FISH) experiment. ( B ) Concentration distribution of SYTO RNASelect (3077 cells for each population from five biological replicates) and EUB338 (1254 cells for each population from three biological replicates) in 1N and multi-N cells. ( C ) The average 1N/multi-N SYTO RNASelect and EUB338 concentration ratio (gray bar) calculated from five and three biological replicates (white circles), respectively. ( D ) RNASelect and EUB338 concentration ratios as functions of cell area (mean ± SD from five and three biological replicates, respectively). Single exponential decay functions were fitted to the average ratios ( R 2 >97%) for each indicated reporter. All concentration comparisons or ratio calculations were performed for equal numbers of 1N and multi-N cells and overlapping cell area distributions (see Materials and methods and ).

Article Snippet: Fluorescence emission was collected during a 100 or 200 ms exposure time provided by a Spectra III Light Engine LED excitation source (Lumencor): mCherry—594 nm excitation, DAPI/FITC/TxRed filter cube (polychroic FF-409/493/596-Di02, triple-pass emitter FF-1-432/523/702-25), GFP and SYTO RNASelect—488 nm excitation, DAPI/FITC/TxRed polychroic filter cube, and an ET525/50M emission filter; YFP—514 nm excitation, CFP/YFP/mCherry filter cube (polychroic FF-459/526/596-Di01, triple-pass emitter FF-1-475/543/702-25), and a FF02-525/40-25 emission filter.

Techniques: CRISPR, Staining, Fluorescence, In Situ Hybridization, Concentration Assay