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

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

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

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

Journal: Cell Death Discovery

Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

doi: 10.1038/s41420-026-03122-x

Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

Article Snippet: Primary human small airway epithelial cells (HSAECs) (PCS-301-010) were obtained from ATCC and cultured in Airway Epithelial Cell Basal Medium (ATCC) containing a Bronchial Epithelial Cell Growth Kit (ATCC) and penicillin-streptomycin (100 U/mL, 100 μg/mL).

Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker

Journal: Antioxidants

Article Title: Nrp1 Signaling Reprograms Glutathione Metabolism to Drive Mitochondrial Dysfunction in Severe Asthma

doi: 10.3390/antiox15040463

Figure Lengend Snippet: Effect of targeting the GSH metabolism on TDI-HSA-treated HAECs. ( A – C ) Detection of cytotoxicity by CCK8 of different groups. ( D ) Measurement of the LDH release rate of different groups. ( E ) MitoSOX and MDC stained HAECs of different groups. Representative H&E-stained HAECs of different groups at 400× original magnification. ( F , G ) Analysis of oxygen consumption rate (OCR) by Seahorse assay. ( H , I ) qPCR analysis of the mRNA expression of TSLP, IL-25, IL-33, E-cadherin, ZO-1 and occludin of the HAECs. ( J ) Relative permeability of HAECs detected by FITC-dextran. GSTO-IN-2 (20 μM), L-glutathione (1 mM). PBS was used as the vehicle (control). * p < 0.05, ** p < 0.005, *** p < 0.0005, ns > 0.05, n = 5.

Article Snippet: Primary human airway epithelial cells (HAECs) were obtained from ATCC.

Techniques: Staining, Expressing, Permeability, Control

Journal: Antioxidants

Article Title: Nrp1 Signaling Reprograms Glutathione Metabolism to Drive Mitochondrial Dysfunction in Severe Asthma

doi: 10.3390/antiox15040463

Figure Lengend Snippet: TDI inhibits SLC25A39 to induce GSH transport and mitochondrial dysfunction. ( A ) Volcano plot of untargeted metabolomics of the mitochondria extracted from healthy and TDI-exposed mice. ( B ) Heat map analysis of oxidized glutathione, glutathione, glutamate, and L-glutamic acid of untargeted metabolomics. ( C ) Construction diagram of the SLC25A39 overexpression plasmid. Asterisk (*) indicates a restriction enzyme site that can be recognized and cleaved by both BspDI and ClaI. ( D ) Measurement of mitochondrial GSH. ( E ) qPCR analysis of the mRNA expression of SLC25A39 in the HAECs. ( F ) Measurement of LDH release rate in the HAECs. ( G ) Detection of cytotoxicity by CCK8 of different groups. ( H ) MDC and MitoSox stained HAECs of different groups. PBS was used as the vehicle (control). * p < 0.05, ** p < 0.005, *** p < 0.0005, ns > 0.05.

Article Snippet: Primary human airway epithelial cells (HAECs) were obtained from ATCC.

Techniques: Over Expression, Plasmid Preparation, Expressing, Staining, Control

Journal: Antioxidants

Article Title: Nrp1 Signaling Reprograms Glutathione Metabolism to Drive Mitochondrial Dysfunction in Severe Asthma

doi: 10.3390/antiox15040463

Figure Lengend Snippet: Effects of targeting the b1 domain of Nrp1 on TDI-HSA-treated primary human airway epithelial cells. ( A , B ) Detection of cytotoxicity by CCK8 of different groups. ( C ) Measurement of the LDH release rate of different groups. ( D ) MitoSOX and MDC stained primary human airway epithelial cells (HAECs) of different groups. ( E , F ) Analysis of OCR by Seahorse assay. ( G ) Representative H&E-stained HAECs of different groups at 400× original magnification. ( H , I ) qPCR analysis of the mRNA expression of TSLP, IL-25, IL-33, E-cadherin, ZO-1 and occludin of the HAECs. ( J ) Relative permeability of HAECs detected by FITC-dextran. EG00229 (5 μM), olopatadine (0.1 mM), dabrafenib (0.5 μM). PBS was used as the vehicle (control). * p < 0.05, ** p < 0.005, *** p < 0.0005, ns > 0.05.

Article Snippet: Primary human airway epithelial cells (HAECs) were obtained from ATCC.

Techniques: Staining, Expressing, Permeability, Control