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Infection and invasion of SkinEthic™ HVE cells by N. gonorrhoeae (a) CLSM z-stack images (x63 objective magnification, zoom setting 3) of the N. gonorrhoeae-infected HVE cells (20 μm) with N. gonorrhoeae pEG2 (expressing sfGFP) shown in green (exc: 488 nm, em: 519 nm) and the nuclei of the <t>rHVE</t> cells in blue (exc: 405 nm, em: 449 nm). Additional supporting images can be found in Biomass of N. gonorrhoeae growth (green channel) quantified using the COMSTAT 2.1 software [ , ]. (d) Quantification of the depth of N. gonorrhoeae infection on the y-axis (green channel, highest to lowest point observed) relative to the depth of remaining HVE cells on the y-axis (blue) in the same field-of-view (measurements in ) as a measure of relative tissue invasion depth. Points in the bar graphs are the mean of values from the five fields-of-view of three biological replicates, with two 20 μm sections each (total 30 field-of-views). Error bars represent the standard error of the mean. Significance values are given as ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. Comparisons which showed no significant differences (p > 0.05) are not indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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Infection and invasion of SkinEthic™ HVE cells by N. gonorrhoeae (a) CLSM z-stack images (x63 objective magnification, zoom setting 3) of the N. gonorrhoeae-infected HVE cells (20 μm) with N. gonorrhoeae pEG2 (expressing sfGFP) shown in green (exc: 488 nm, em: 519 nm) and the nuclei of the <t>rHVE</t> cells in blue (exc: 405 nm, em: 449 nm). Additional supporting images can be found in Biomass of N. gonorrhoeae growth (green channel) quantified using the COMSTAT 2.1 software [ , ]. (d) Quantification of the depth of N. gonorrhoeae infection on the y-axis (green channel, highest to lowest point observed) relative to the depth of remaining HVE cells on the y-axis (blue) in the same field-of-view (measurements in ) as a measure of relative tissue invasion depth. Points in the bar graphs are the mean of values from the five fields-of-view of three biological replicates, with two 20 μm sections each (total 30 field-of-views). Error bars represent the standard error of the mean. Significance values are given as ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. Comparisons which showed no significant differences (p > 0.05) are not indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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Infection and invasion of SkinEthic™ HVE cells by N. gonorrhoeae (a) CLSM z-stack images (x63 objective magnification, zoom setting 3) of the N. gonorrhoeae-infected HVE cells (20 μm) with N. gonorrhoeae pEG2 (expressing sfGFP) shown in green (exc: 488 nm, em: 519 nm) and the nuclei of the <t>rHVE</t> cells in blue (exc: 405 nm, em: 449 nm). Additional supporting images can be found in Biomass of N. gonorrhoeae growth (green channel) quantified using the COMSTAT 2.1 software [ , ]. (d) Quantification of the depth of N. gonorrhoeae infection on the y-axis (green channel, highest to lowest point observed) relative to the depth of remaining HVE cells on the y-axis (blue) in the same field-of-view (measurements in ) as a measure of relative tissue invasion depth. Points in the bar graphs are the mean of values from the five fields-of-view of three biological replicates, with two 20 μm sections each (total 30 field-of-views). Error bars represent the standard error of the mean. Significance values are given as ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. Comparisons which showed no significant differences (p > 0.05) are not indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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Infection and invasion of SkinEthic™ HVE cells by N. gonorrhoeae (a) CLSM z-stack images (x63 objective magnification, zoom setting 3) of the N. gonorrhoeae-infected HVE cells (20 μm) with N. gonorrhoeae pEG2 (expressing sfGFP) shown in green (exc: 488 nm, em: 519 nm) and the nuclei of the rHVE cells in blue (exc: 405 nm, em: 449 nm). Additional supporting images can be found in Biomass of N. gonorrhoeae growth (green channel) quantified using the COMSTAT 2.1 software [ , ]. (d) Quantification of the depth of N. gonorrhoeae infection on the y-axis (green channel, highest to lowest point observed) relative to the depth of remaining HVE cells on the y-axis (blue) in the same field-of-view (measurements in ) as a measure of relative tissue invasion depth. Points in the bar graphs are the mean of values from the five fields-of-view of three biological replicates, with two 20 μm sections each (total 30 field-of-views). Error bars represent the standard error of the mean. Significance values are given as ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. Comparisons which showed no significant differences (p > 0.05) are not indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Journal: Biofilm

Article Title: Influence of the ATP-dependent DNA ligase, Lig E, on Neisseria gonorrhoeae microcolony and biofilm formation

doi: 10.1016/j.bioflm.2025.100292

Figure Lengend Snippet: Infection and invasion of SkinEthic™ HVE cells by N. gonorrhoeae (a) CLSM z-stack images (x63 objective magnification, zoom setting 3) of the N. gonorrhoeae-infected HVE cells (20 μm) with N. gonorrhoeae pEG2 (expressing sfGFP) shown in green (exc: 488 nm, em: 519 nm) and the nuclei of the rHVE cells in blue (exc: 405 nm, em: 449 nm). Additional supporting images can be found in Biomass of N. gonorrhoeae growth (green channel) quantified using the COMSTAT 2.1 software [ , ]. (d) Quantification of the depth of N. gonorrhoeae infection on the y-axis (green channel, highest to lowest point observed) relative to the depth of remaining HVE cells on the y-axis (blue) in the same field-of-view (measurements in ) as a measure of relative tissue invasion depth. Points in the bar graphs are the mean of values from the five fields-of-view of three biological replicates, with two 20 μm sections each (total 30 field-of-views). Error bars represent the standard error of the mean. Significance values are given as ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. Comparisons which showed no significant differences (p > 0.05) are not indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: We also used commercially available 3-dimensional (3-D) reconstituted human vaginal epithelium (rHVE) (SkinEthic Laboratories) in lieu of traditional 2-dimensional (2-D) cell lines for host-cell assays.

Techniques: Infection, Expressing, Software

Quantification of lactate dehydrogenase (LDH) release in the supernatant after N. gonorrhoeae infection of rHVE tissue. Relative LDH activity was obtained via absorbance at 490 nm relative to the absorbance of blank medium and corrected for by the background absorbance at 680 nm (raw data in ). Points (data in ) are the mean of values from three technical replicates of each biological replicate and error bars represent the standard error of the mean. Significance values are given as ∗ p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 Comparisons which showed no significant differences (p > 0.05) are not indicated.

Journal: Biofilm

Article Title: Influence of the ATP-dependent DNA ligase, Lig E, on Neisseria gonorrhoeae microcolony and biofilm formation

doi: 10.1016/j.bioflm.2025.100292

Figure Lengend Snippet: Quantification of lactate dehydrogenase (LDH) release in the supernatant after N. gonorrhoeae infection of rHVE tissue. Relative LDH activity was obtained via absorbance at 490 nm relative to the absorbance of blank medium and corrected for by the background absorbance at 680 nm (raw data in ). Points (data in ) are the mean of values from three technical replicates of each biological replicate and error bars represent the standard error of the mean. Significance values are given as ∗ p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 Comparisons which showed no significant differences (p > 0.05) are not indicated.

Article Snippet: We also used commercially available 3-dimensional (3-D) reconstituted human vaginal epithelium (rHVE) (SkinEthic Laboratories) in lieu of traditional 2-dimensional (2-D) cell lines for host-cell assays.

Techniques: Infection, Activity Assay