Journal: Archives of Toxicology

Article Title: Involvement of necroptosis in the selective toxicity of the natural compound (±) gossypol on squamous skin cancer cells in vitro

doi: 10.1007/s00204-023-03516-1

Figure Lengend Snippet: Effect of ABT-199 on cell viability of SCL-1 and NHEK. A Chemical structure of ABT-199/venetoclax. To determine the effect of ABT-199 on cell viability, SCL-1 carcinoma cells (B) and keratinocytes (NHEK) (C) were treated with different concentrations of ABT-199 for 96 h. Cell viability was measured by MTT assay. Mock-treated control was set at 100%. Data represent means ± SEM of at least three independent experiments ( n ≥ 3). One-way ANOVA with Dunnett’s multiple comparison test was used for the determination of statistical significance compared to mock-treated control; ** p < 0.01, *** p < 0.001

Article Snippet: For treatment, SCL-1 and A375 were incubated in high glucose (4500 mg/L) DMEM without FBS, while NHEK were grown in keratinocyte basal medium (C-20211, PromoCell).

Techniques: MTT Assay, Control, Comparison

Journal: Archives of Toxicology

Article Title: Involvement of necroptosis in the selective toxicity of the natural compound (±) gossypol on squamous skin cancer cells in vitro

doi: 10.1007/s00204-023-03516-1

Figure Lengend Snippet: Selective effect of GP on cell viability of skin cells. A Chemical structure of gossypol (GP). To examine the effect of GP on cell viability, SCL-1 carcinoma cells and keratinocytes (NHEK) were treated with different concentrations of GP for 96 h or mock treated (0) and cell viability was measured by MTT (B) and SRB (C) assay. Mock-treated controls were set at 100%. Data represent means ± SEM of at least three independent experiments ( n ≥ 3). Student’s t test was used for the determination of statistical significance; ** p < 0.01, *** p < 0.001. D, E IC 50 values were calculated by non-linear curve fit analysis (GraphPad Prism 5) based on MTT (D) and SRB assay (E)

Article Snippet: For treatment, SCL-1 and A375 were incubated in high glucose (4500 mg/L) DMEM without FBS, while NHEK were grown in keratinocyte basal medium (C-20211, PromoCell).

Techniques: Sulforhodamine B Assay

Journal: Archives of Toxicology

Article Title: Involvement of necroptosis in the selective toxicity of the natural compound (±) gossypol on squamous skin cancer cells in vitro

doi: 10.1007/s00204-023-03516-1

Figure Lengend Snippet: Intracellular amount of GP in SCL-1 cells and keratinocytes (NHEK). Cells were harvested after treatment with 5 µM GP for 0.25, 1, and 2 h and analyzed by HPLC. The amount of intracellular GP was related to the respective protein level. Experiments were performed in triplicate ( n = 3)

Article Snippet: For treatment, SCL-1 and A375 were incubated in high glucose (4500 mg/L) DMEM without FBS, while NHEK were grown in keratinocyte basal medium (C-20211, PromoCell).

Techniques:

Journal: Archives of Toxicology

Article Title: Involvement of necroptosis in the selective toxicity of the natural compound (±) gossypol on squamous skin cancer cells in vitro

doi: 10.1007/s00204-023-03516-1

Figure Lengend Snippet: Mitochondrial respiration of GP-treated normal keratinocytes (NHEK). A After treatment with different concentrations of GP or mock treated for 1 h, the oxygen consumption rate (OCR) was measured after successive injection of oligomycin (Oligo), FCCP, and rotenone/antiymcin A (Rot/AA) by Seahorse XF Analyzer. Representative curves are depicted. B–D Based on the OCR in response to these mitochondrial stressors, ATP production (B) , spare respiratory capacity (C), and proton leak (D) were calculated. The values of untreated cells were set at 100%. Data represent means ± SEM of three independent experiments ( n = 3). One-way ANOVA with Dunnett’s multiple comparison test was used for the determination of statistical significance between mock treated and GP-treated cells; * p < 0.01

Article Snippet: For treatment, SCL-1 and A375 were incubated in high glucose (4500 mg/L) DMEM without FBS, while NHEK were grown in keratinocyte basal medium (C-20211, PromoCell).

Techniques: Injection, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Radiofrequency Treatment Attenuates Age-Related Changes in Dermal–Epidermal Junctions of Animal Skin

doi: 10.3390/ijms25105178

Figure Lengend Snippet: Expression of HSP47, HSP90, TGF-β, and DEJ proteins following RF treatment in senescent keratinocytes. ( A ) Schematic representation of non-senescent keratinocytes, senescent keratinocytes, and RF treatments administered to senescent keratinocytes. ( B – H ) Proteins were extracted from senescent keratinocytes following RF treatment and subjected to the following assays. ( B – D ) Western blot analysis of HSP47 and HSP90. ( E ) ELISA results for TGF-β. ( F , G ) Western blot analysis of collagen XVII. ( H ) ELISA results for collagen IV. Data are presented as mean ± SD of three independent experiments. **, p < 0.01 vs. first bar; $$, p < 0.01 vs. second bar (Mann–Whitney U test). CON, control; DEJ, dermal–epidermal junction; ELISA, enzyme-linked immunosorbent assay; GM, growth medium; HSP47, heat shock protein 47; HSP90, heat shock protein 90; MW, molecular weight; PBS, phosphate-buffered saline; RF, radiofrequency; SD, standard deviation; SnCs, senescent; TGF-β, transforming growth factor beta.

Article Snippet: Primary human epidermal keratinocytes (HEKn; American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in growth medium (GM) composed of dermal cell basal medium (ATCC) supplemented with a keratinocyte growth kit (ATCC).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control, Molecular Weight, Saline, Standard Deviation