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Host cell damage and virulence capacity of mutants in sugar nucleotide biosynthesis. (A) Mutants grown in +/− 25 μg/ml Dox screened for <t>epithelial</t> damage using A-431 cells by LDH assay. The mean LDH released at 24 h post co-incubation is shown for repressed mutants (grown in presence of Dox; blue bars) and No-Dox controls (red bars). Red and blue horizontal lines indicate the mean LDH activity for wild type control (No-Dox) and wild type grown in presence of Dox respectively. Welsh t-test used for statistical analysis; error bars represent standard error of mean; p**** < 0.0001. (B) Survival plots of G. mellonella larvae infected with C. albicans mutants in: (I) GDP-mannose, (II) UDP-glucose and (III) UDP- N -acetylglucosamine biosynthesis in presence (solid lines) and absence (dotted lines) of Dox. No killing or improved survival was observed for a number of repressed mutants. No killing was observed in control larvae injected with equivalent volume of PBS. (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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Host cell damage and virulence capacity of mutants in sugar nucleotide biosynthesis. (A) Mutants grown in +/− 25 μg/ml Dox screened for <t>epithelial</t> damage using A-431 cells by LDH assay. The mean LDH released at 24 h post co-incubation is shown for repressed mutants (grown in presence of Dox; blue bars) and No-Dox controls (red bars). Red and blue horizontal lines indicate the mean LDH activity for wild type control (No-Dox) and wild type grown in presence of Dox respectively. Welsh t-test used for statistical analysis; error bars represent standard error of mean; p**** < 0.0001. (B) Survival plots of G. mellonella larvae infected with C. albicans mutants in: (I) GDP-mannose, (II) UDP-glucose and (III) UDP- N -acetylglucosamine biosynthesis in presence (solid lines) and absence (dotted lines) of Dox. No killing or improved survival was observed for a number of repressed mutants. No killing was observed in control larvae injected with equivalent volume of PBS. (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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Barplots showing percentage of viability of <t>S9</t> <t>epithelial</t> cells following administration of different concentrations of herbicolin A (HrbA) and B (HrbB) (1: 0.5, 2: 2.5, 3: 10, and 4: 50 μg/mL). NC: negative control represents no treatment. PC: Positive control represents 1% Triton X. Box-and-whisker plots show the median (horizontal line), interquartile range (box), and minimum and maximum values (whiskers). Data are shown as individual values with group means. Statistical significance was assessed using one-way ANOVA followed by Holm-corrected pairwise Student’s t -tests. The equation shown at the top of the plot reveals the statistical model used for comparisons. The values shown at the bottom of the plot indicate the ANOVA results, including the F-statistic with degrees of freedom, the associated p-value, and the effect size with 95% confidence interval.
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Barplots showing percentage of viability of <t>S9</t> <t>epithelial</t> cells following administration of different concentrations of herbicolin A (HrbA) and B (HrbB) (1: 0.5, 2: 2.5, 3: 10, and 4: 50 μg/mL). NC: negative control represents no treatment. PC: Positive control represents 1% Triton X. Box-and-whisker plots show the median (horizontal line), interquartile range (box), and minimum and maximum values (whiskers). Data are shown as individual values with group means. Statistical significance was assessed using one-way ANOVA followed by Holm-corrected pairwise Student’s t -tests. The equation shown at the top of the plot reveals the statistical model used for comparisons. The values shown at the bottom of the plot indicate the ANOVA results, including the F-statistic with degrees of freedom, the associated p-value, and the effect size with 95% confidence interval.
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ATCC human normal pancreatic ductal epithelial cells hpne
The expression of EN2 in <t>HPNE,</t> <t>pancreatic</t> cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.
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Host cell damage and virulence capacity of mutants in sugar nucleotide biosynthesis. (A) Mutants grown in +/− 25 μg/ml Dox screened for epithelial damage using A-431 cells by LDH assay. The mean LDH released at 24 h post co-incubation is shown for repressed mutants (grown in presence of Dox; blue bars) and No-Dox controls (red bars). Red and blue horizontal lines indicate the mean LDH activity for wild type control (No-Dox) and wild type grown in presence of Dox respectively. Welsh t-test used for statistical analysis; error bars represent standard error of mean; p**** < 0.0001. (B) Survival plots of G. mellonella larvae infected with C. albicans mutants in: (I) GDP-mannose, (II) UDP-glucose and (III) UDP- N -acetylglucosamine biosynthesis in presence (solid lines) and absence (dotted lines) of Dox. No killing or improved survival was observed for a number of repressed mutants. No killing was observed in control larvae injected with equivalent volume of PBS. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Journal: The Cell Surface

Article Title: Compromising UPD-sugar nucleotide biosynthesis attenuates Candida albicans viability, virulence and drug sensitivity ☆

doi: 10.1016/j.tcsw.2026.100170

Figure Lengend Snippet: Host cell damage and virulence capacity of mutants in sugar nucleotide biosynthesis. (A) Mutants grown in +/− 25 μg/ml Dox screened for epithelial damage using A-431 cells by LDH assay. The mean LDH released at 24 h post co-incubation is shown for repressed mutants (grown in presence of Dox; blue bars) and No-Dox controls (red bars). Red and blue horizontal lines indicate the mean LDH activity for wild type control (No-Dox) and wild type grown in presence of Dox respectively. Welsh t-test used for statistical analysis; error bars represent standard error of mean; p**** < 0.0001. (B) Survival plots of G. mellonella larvae infected with C. albicans mutants in: (I) GDP-mannose, (II) UDP-glucose and (III) UDP- N -acetylglucosamine biosynthesis in presence (solid lines) and absence (dotted lines) of Dox. No killing or improved survival was observed for a number of repressed mutants. No killing was observed in control larvae injected with equivalent volume of PBS. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human epithelial cells derived from a vulvar squamous cell carcinoma (A-431 cell line; ATCC No.: CRL-1555) were cultured and maintained in DMEM medium supplemented with 10% ( v /v) heat inactivated foetal calf serum, 5% penicillin and 5% streptomycin.

Techniques: Lactate Dehydrogenase Assay, Incubation, Activity Assay, Control, Infection, Injection

Barplots showing percentage of viability of S9 epithelial cells following administration of different concentrations of herbicolin A (HrbA) and B (HrbB) (1: 0.5, 2: 2.5, 3: 10, and 4: 50 μg/mL). NC: negative control represents no treatment. PC: Positive control represents 1% Triton X. Box-and-whisker plots show the median (horizontal line), interquartile range (box), and minimum and maximum values (whiskers). Data are shown as individual values with group means. Statistical significance was assessed using one-way ANOVA followed by Holm-corrected pairwise Student’s t -tests. The equation shown at the top of the plot reveals the statistical model used for comparisons. The values shown at the bottom of the plot indicate the ANOVA results, including the F-statistic with degrees of freedom, the associated p-value, and the effect size with 95% confidence interval.

Journal: bioRxiv

Article Title: Herbicolin A, an antifungal lipopeptide produced by Pantoea agglomerans APC 4211 is a promising biocontrol agent against food spoilage fungi

doi: 10.64898/2026.05.21.726617

Figure Lengend Snippet: Barplots showing percentage of viability of S9 epithelial cells following administration of different concentrations of herbicolin A (HrbA) and B (HrbB) (1: 0.5, 2: 2.5, 3: 10, and 4: 50 μg/mL). NC: negative control represents no treatment. PC: Positive control represents 1% Triton X. Box-and-whisker plots show the median (horizontal line), interquartile range (box), and minimum and maximum values (whiskers). Data are shown as individual values with group means. Statistical significance was assessed using one-way ANOVA followed by Holm-corrected pairwise Student’s t -tests. The equation shown at the top of the plot reveals the statistical model used for comparisons. The values shown at the bottom of the plot indicate the ANOVA results, including the F-statistic with degrees of freedom, the associated p-value, and the effect size with 95% confidence interval.

Article Snippet: Human airway epithelial cells (S9) were obtained from the American Type Culture Collection (LGC Promochem, Wesel, Germany) (no. CRL-2778).

Techniques: Negative Control, Positive Control, Whisker Assay

The expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: The expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.

Article Snippet: Human pancreatic cancer cell lines (PANC‐1 and AsPC‐1) and human normal pancreatic ductal epithelial cells (HPNE) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Isolation, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry

Overexpression of EN2 in HPNE cells induces cellular transformation and stemness. (A and B) HPNE cells were stably transduced with lentiviral particles expressing either empty vector or EN2 cDNA. EN2 expression was measured by immunocytochemistry and qRT‐PCR. Blue colour = nuclei; Green colour = EN2. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). (C), Spheroid formation in suspension. Spheroid formation of HPNE/Empty Vector and HPNE/EN2 cDNA cells was measured. Spheroids in suspensions were photographed (left) and counted (right). Data represent mean ( n = 4) ± SD. # = significantly different between groups ( p < 0.05). (D), Expression of stem cell markers. RNA was isolated, and expression of stem cell markers (CD24, CD44, CD133 and LGR5) was measured by qRT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1. (E), Expression of pluripotency‐maintaining factors. RNA was isolated, and the expression of pluripotency‐maintaining factors (Oct4, Sox2, cMyc and KLF4) was measured by qRT‐PCR analysis. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1.

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: Overexpression of EN2 in HPNE cells induces cellular transformation and stemness. (A and B) HPNE cells were stably transduced with lentiviral particles expressing either empty vector or EN2 cDNA. EN2 expression was measured by immunocytochemistry and qRT‐PCR. Blue colour = nuclei; Green colour = EN2. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). (C), Spheroid formation in suspension. Spheroid formation of HPNE/Empty Vector and HPNE/EN2 cDNA cells was measured. Spheroids in suspensions were photographed (left) and counted (right). Data represent mean ( n = 4) ± SD. # = significantly different between groups ( p < 0.05). (D), Expression of stem cell markers. RNA was isolated, and expression of stem cell markers (CD24, CD44, CD133 and LGR5) was measured by qRT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1. (E), Expression of pluripotency‐maintaining factors. RNA was isolated, and the expression of pluripotency‐maintaining factors (Oct4, Sox2, cMyc and KLF4) was measured by qRT‐PCR analysis. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1.

Article Snippet: Human pancreatic cancer cell lines (PANC‐1 and AsPC‐1) and human normal pancreatic ductal epithelial cells (HPNE) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Over Expression, Transformation Assay, Stable Transfection, Transduction, Expressing, Plasmid Preparation, Immunocytochemistry, Quantitative RT-PCR, Suspension, Isolation, Control, Gene Expression

Overexpression of EN2 in HPNE cells enhances cell motility and modulates expression of EMT‐related genes. (A) Cell Motility. HPNE cells were stably transduced with lentiviral particles expressing either empty vector or EN2 cDNA. (B) Expression of EMT‐related genes. RNA was isolated, and the EMT‐related genes (E‐cadherin, N‐cadherin, Snail, Slug, and Zeb1) were measured by qRT‐PCR analysis. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1.

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: Overexpression of EN2 in HPNE cells enhances cell motility and modulates expression of EMT‐related genes. (A) Cell Motility. HPNE cells were stably transduced with lentiviral particles expressing either empty vector or EN2 cDNA. (B) Expression of EMT‐related genes. RNA was isolated, and the EMT‐related genes (E‐cadherin, N‐cadherin, Snail, Slug, and Zeb1) were measured by qRT‐PCR analysis. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( p < 0.05). Gene expression of Empty Vector was normalised to 1.

Article Snippet: Human pancreatic cancer cell lines (PANC‐1 and AsPC‐1) and human normal pancreatic ductal epithelial cells (HPNE) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Over Expression, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Isolation, Quantitative RT-PCR, Control, Gene Expression