Journal: Toxicology and applied pharmacology

Article Title: Monoubiquitinated γ-H2AX: abundant product and specific biomarker for non-apoptotic DNA double-strand breaks

doi: 10.1016/j.taap.2018.07.007

Figure Lengend Snippet: Panels A-C used antibodies #1. (A) Impact of gel polyacrylamide percentage and transfer conditions to PVDF membranes. Treatments of H460 cells: 6 h with 3 mM hydroxyurea (HU), 6 or 16 h with 5 μM aphidicolin (Aphi) or 1 h with 5 μM bleomycin (Bleo). All γ-H2AX images had 5 s film exposures. (B) Ethanol effect in semi-dry transfer. IMR90 cells were treated with 40 μM etoposide (Eto) for 1 h. (C) Comparison of wet (pH 8.2 and pH 8.8) and semi-dry transfers using 12% ethanol-containing buffers (10 s film exposures for all γ-H2AX images). IMR90 cells were treated with 40 μM etoposide for 1 h. (D) Effectiveness of different antibodies using semi-dry transfer with 12% ethanol. Normal human keratinocytes were treated for 1 h with 30 μM etoposide or 0.5 μM bleomycin. Antibodies #1–3: 1:1000 dilutions and 5 s exposures, antibody #4: 1:1000 dilution and 60 s exposure. (E) Comparison of antibodies #1 and #5 (both at 1:1000 dilutions) in H460 cells treated with bleomycin for 1 h. (F) γ-H2AX blot (Ab#1) in control and H2AX-depleted IMR90 cells treated with 100 μM etoposide for 1h.

Article Snippet: IMR90, WI38, normal human neonatal keratinocytes (PCS-200–010), H460, A549, U2OS, mouse embryonic fibroblasts (MEFs, SCRC-1040), telomerase-immortalized human bronchial epithelial HBEC3 (CRL-4051), Daudi, HCT116, Ramos, Raji and CA46 were obtained from ATCC.

Techniques: Comparison, Control

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: International Journal of Molecular Sciences

Article Title: Cell-Penetrating Delivery of Nitric Oxide by Biocompatible Dinitrosyl Iron Complex and Its Dermato-Physiological Implications

doi: 10.3390/ijms221810101

Figure Lengend Snippet: Cell viability assay of the ( a ) mouse skin melanoma (B16-F10) and ( b ) human skin keratinocyte cells (HaCaT) cells, respectively, treated with different concentrations of DNIC-2 for 24 h **** p < 0.001 compared to the group without treatment of DNIC-2 . ( c ) Cell viability assay of the reconstructed human epidermis (RhE) model treated with PBS, 5% SDS, and 50 μM of DNIC-2 , respectively. ( d ) Cell viability assay of the reconstructed human cornea-like epithelium model treated with DPBS, methyl acetate, and 50 μM of DNIC-2 , respectively.

Article Snippet: CCD-966Sk human skin fibroblasts and B16-F10 mouse skin melanoma cells were purchased from Bioresource Collection and Research Center, Food Industry Research and Development Institute (Hsinchu, Taiwan), whereas human skin keratinocyte cell line, HaCaT, was purchased from Elabscience Biotechnology Inc. (Elabscience ® EP-CL-0090, Houston, TX, USA).

Techniques: Viability Assay

Journal: Frontiers in Pharmacology

Article Title: Syzygium aqueum : A Polyphenol- Rich Leaf Extract Exhibits Antioxidant, Hepatoprotective, Pain-Killing and Anti-inflammatory Activities in Animal Models

doi: 10.3389/fphar.2018.00566

Figure Lengend Snippet: Effect of Syzygium aqueum extracts on human keratinocytes. (A) Biocompatibility of S. aqueum extract in dose (25–200 μg/mL) and time (24–48 h) manner. Survival rate of the cells was assessed by the MTT assay. (B) ROS production in cells treated or untreated with theextract (100 μg/mL) and then irradiated by 100 J/cm 2 UVA (+) or not (–). (C) Total GSH levels determined by DTNB assay. Results expressed as mean ± SD of three assays. Significant ∗ p < 0.01; ∗∗ p < 0.001 compared to untreated control. Analysis performed by two-way ANOVA followed by Bonferroni’s multiple comparisons test. (D) Immunoblotting bands of the oxidative stress activated protein, p38. An equal amount (100 μg) of total lysate from each sample was resolved by SDS-PAGE with GAPDH as a control.

Article Snippet: Human epidermal keratinocytes (HaCaT), provided by Innoprot (Biscay, Spain), were cultured as described in .

Techniques: MTT Assay, Irradiation, DTNB Assay, Western Blot, SDS Page

Journal: Arthritis Research & Therapy

Article Title: Synergistic role of c-Myc and ERK1/2 in the mitogenic response to TGFβ-1 in cultured rat nucleus pulposus cells

doi: 10.1186/ar2567

Figure Lengend Snippet: Effect of transforming growth factor β1 (TGFβ1) treatment on mRNA expression in different cell types (a), Cells were treated with or without 5 ng/mL TGFβ1 for 24 h . The expression of c-myc in nucleus pulposus cells (NP), in articular chondrocytes (AC) and keratinocytes (KT) are presented. The expression of p15 , p21 and p27 in NP was also determined. Time course of c-myc expression in NP treated with 5 ng/mL TGFβ1 (b). The graph shows the relative intensities of c-myc bands normalized for β-actin levels by densitographic analysis. Incubation for 24 h with medium containing various concentrations of fetal bovine serum (FBS) did not alter the level of c-myc expression in NP (c). The reverse transcription-polymerase chain reaction (RT-PCR) was performed on total RNA extracted from the cells. β-actin was used as an internal control.

Article Snippet: Cryopreserved primary passage rat epidermal keratinocytes were obtained from Cell Applications Inc. (San Diego, CA, USA) and maintained in growth medium (Cell Applications Inc.).

Techniques: Expressing, Incubation, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Control