Journal: iScience

Article Title: Topical application of urolithin A activates mitophagy and enhances mitochondrial function to promote skin longevity

doi: 10.1016/j.isci.2026.117646

Figure Lengend Snippet: UA beneficial impact on human keratinocytes and reconstituted human epidermis (A) Mitochondrial respiration (basal and maximal) assessed as oxygen consumption rate (OCR) in human dermal keratinocytes after 72 h of differentiation from young donor treated with DMSO for 24 h or the indicated dose of UA. ∗ p < 0.05 and ∗∗∗ p < 0.001 after one-way ANOVA. Error bars represent mean ± SEM. (B) TOMM20 immunostaining of skin epidermis in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as antibody staining signal over imaging field. ∗ p < 0.05 and ∗∗∗∗ p < 0.0001 after unpaired t test one-way ANOVA. Error bars represent mean ± SEM. Scale bars, 50 μm. (C) Relative gene expression of PINK1 and PARK2 in reconstructed human epidermis (RHE). RHE tissues were moisturized for 24 h with or without 1% Mitopure, and relative expression levels of PINK1 and PARK2 were quantified. Statistical analysis was performed using one-way ANOVA (∗ p < 0.05; ∗∗ p < 0.01). Error bars represent mean ± SEM. (D) Interleukin 6 levels normalized to cell viability in keratinocytes. Keratinocytes were stimulated with a pro-inflammatory cytokine mix for 24 h then treated for an additional 24 h with either DMSO or 1 μM UA. IL-6 secretion was quantified and normalized to cell viability. Statistical analysis was performed using one-way ANOVA (∗∗∗∗ p < 0.0001). Error bars represent mean ± SEM. (E) p16 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as antibody staining signal over imaging field. ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm. (F) Ki67 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as number of Ki67 positive cells normalized over μm of dermal epidermal junction (count/μm) ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm. (G) Keratin 10 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as fluorescence intensity over epidermis area. ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm.

Article Snippet: Human: Normal Human Epidermal Keratinocytes (NHEK) , PromoCell , Cat#C-12003; Lot#479Z011.1.

Techniques: Immunostaining, Staining, Imaging, Gene Expression, Expressing, Analysis, Fluorescence

Journal: iScience

Article Title: Topical application of urolithin A activates mitophagy and enhances mitochondrial function to promote skin longevity

doi: 10.1016/j.isci.2026.117646

Figure Lengend Snippet:

Article Snippet: Human: Normal Human Epidermal Keratinocytes (NHEK) , PromoCell , Cat#C-12003; Lot#479Z011.1.

Techniques: Binding Assay, Ex Vivo, Recombinant, Metals, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Cell Viability Assay, Staining, DC Protein Assay, Gene Expression, Glass, Software, Imaging, Microscopy, Real-time Polymerase Chain Reaction

Journal:

Article Title: A Homozygous Missense Mutation in TGM5 Abolishes Epidermal Transglutaminase 5 Activity and Causes Acral Peeling Skin Syndrome

doi:

Figure Lengend Snippet: Biochemical studies of TG5 mutant forms. NHEK (a) and HEK-293 (b) cells were transfected for 48 h with the use of wild-type TG5 and the three mutant forms indicated. Total TG5 activities were measured for insoluble and fractions and standardized for transfection efficiency (GFP). The control vector indicates the endogenous TG5 activity (cells transfected only with the GFP control vector). The data obtained are reported as percentages of TG activity, with 100% corresponding to the activity for cells transfected with the wild-type construct. The data presented are the averages of two independent experiments. c, View of the entire TG5 model with the four main structural domains. The mutations described here are located in the N-terminal domain at the interface between the N-terminal domain and the catalytic domain. d, Effects of the mutations in a local environment. The G113C mutation, close to the active site, would probably cause a local change in the main protein structure. The poorly conserved residue T109 is present on the surface and is, therefore, unlikely to be functionally critical. Green indicates the mutated residues, and red represents wild-type TG5 residues.

Article Snippet: Cell Culture and Transfection Cryopreserved NHEK cells obtained from BioWhittaker were grown in calf skin collagen–coated dishes (Sigma Chemical) in serum-free keratinocyte medium (BioWhittaker) at 0.05 mM Ca 2+ and supplemented with Single-Quots (BioWhittaker) containing 7.5 μg/ml bovine pituitary extract, 0.5 mg/ml insulin, 0.5 mg/ml hydrocortisone, and 0.1 μg/ml hEGF.

Techniques: Mutagenesis, Transfection, Control, Plasmid Preparation, Activity Assay, Construct, Residue

Journal: Pharmaceutics

Article Title: Tissue Engineered Human Skin Equivalents

doi: 10.3390/pharmaceutics4010026

Figure Lengend Snippet: Summary of commercially available human skin equivalents.

Article Snippet: Epiderm , MatTek , , Neonatal human-derived epidermal keratinocytes (NHEK) cultured to form a multi-layered, highly differentiated model of the human epidermis.

Techniques: Ex Vivo, Cell Culture, In Vitro, Functional Assay

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Commercially available psoriasis 3D skin models for in vitro applications

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: In Vitro, Ex Vivo, Membrane

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Overview of applications using psoriasis(-induced) skin substitutes

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: Membrane, Expressing, Migration, In Vivo

Journal: Experimental Biology and Medicine

Article Title: In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research

doi: 10.1177/1535370217710637

Figure Lengend Snippet: Overview of the different types of psoriasis models with pros and cons, and their most current applications

Article Snippet: • MatTek FT , NHEK + PHDF , MatTek cytokine mix , Drug testing: PTTC, calcipotriol, IL-4, coal tar, psoriasin gel, delphinidin , 84 – 88.

Techniques: Drug discovery, Ex Vivo, Migration, Activity Assay, Transgenic Assay, Control

Journal:

Article Title: Development of a Frozen Cell Array as a High-Throughput Approach for Cell-Based Analysis

doi:

Figure Lengend Snippet: Binding of an anti-human Ep-CAM fluorescein-conjugated mAb to various cell lines analyzed by FACS. BKGEs (A2); human colorectal carcinoma cell lines COLO205 (A3), HCT116 (A4), and SW620 (A5); human prostate carcinoma cell lines PC3 (B1) and DU145 (B2); rat prostate tumorigenic cell line NRP154 (B3); HIAECs (B4); HUVECs (B5); lung HMVECs (C1); NHDFs (C2); human CASMCs (C3); neonatal NHEKs (C4); human malignant melanoma cell line A375 (C5); human breast carcinoma cell lines SkBr3 (D1), MCF7 (D2), and BT474 (D3); human rhabdomyosarcoma cell line A673 (D4); human liver carcinoma cell lines HepG2 (D5) and Hep3b (E1); T- and B-cell leukemia cell lines Jurkat (E2) and Bjab (E3), respectively; human lung carcinoma cell line SKMES1 (E4); and human neuroglioma cell line U87MG (E5) were incubated in the absence (thin line) or presence (thick line) of an anti-human Ep-CAM fluorescein-conjugated monoclonal antibody and analyzed by FACS.

Article Snippet: Normal cells included: human iliac artery endothelial cells (HIAECs), human umbilical vein endothelial cells (HUVECs), human microvascular endothelial cells from lung (HMVECs), normal human dermal fibroblasts (NHDFs), human coronary artery smooth muscle cells (CASMCs), and neonatal normal human epidermal keratinocytes (NHEKs) purchased from BioWhittaker/Clonetics (San Diego, CA) and the bovine kidney glomerular endothelial cell (BKGEs) purchased from VEC Technologies, Inc. (Rensselaer, NY, USA).

Techniques: Binding Assay, Incubation

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mechanistic Contribution of Ubiquitous 15-Lipoxygenase-1 Expression Loss in Cancer Cells to Terminal Cell Differentiation Evasion

doi: 10.1158/1940-6207.CAPR-10-0280

Figure Lengend Snippet: 15-LOX-1 and P16 mRNA and protein expression levels in cancer cell lines. A and B, A total of 128 cancer cell lines (Supplementary Table S1) were cultured and processed for 15-LOX-1 (panel A) and p16 (panel B) mRNA by quantitative real-time reverse transcription-polymerase chain reaction. Dots in the dot plots are means of triplicate measurements from each cell line. The relative expression levels were calculated relative to expression of the calibrator sample (differentiated NHBE cells). Solid lines represent the median value for each group. C, 15-LOX-1 relative expression levels in cancer cell lines compared to the level in Caco-2 cells terminally differentiated by sodium butyrate treatment. 15-LOX-1 mRNA measurements are as in panel A but with terminally differentiated Caco-2 cells as the calibrator sample. Dots in the dot plots are means of triplicate measurements from each cell line. The solid line represents the median value for the relative expression levels. D, 15-LOX-1 protein expression in cancer cell lines. Cell lines—including cell lines with 15-LOX-1 mRNA expression levels nearly equal to or greater than the level in differentiated Caco-2 cells or NHEK—were cultured, processed for Western blotting, and probed with 15-LOX-1 antibody. Three repeated experiments yielded similar results.

Article Snippet: We also purchased from MatTek Corporation NHEK cultured in three-dimensional orientation to maintain air-liquid interface and to form a multilayered, highly differentiated structure that highly resembles human epidermis (differentiated NHEK) ( Supplementary Fig. S1C ) ( 24 ).

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot