Journal: Molecules

Article Title: Green Synthesis of Au-Pd Bimetallic Nanoparticles Using Aspalathin and Their Toxicity Study

doi: 10.3390/molecules31050910

Figure Lengend Snippet: Cell viability of SKMEL-1 and HaCAT cells was assessed using the MTT assay during a 24 h exposure to different samples, including GR ( A ), ASP ( B ), GR-Au-PdNPs ( C ), ASP-AuPdNPs ( D ), GR ( E ), ASP ( F ), GR-Au-PdNPs ( G ), and ASP-AuPdNPs ( H ).

Article Snippet: Cell lines, namely SKMEL-1 (human melanoma cell line), HaCAT (human epidermal keratinocyte cell line), KMST-6 (human fibroblast cell line), and HepG2 (liver hepatocellular carcinoma cell line), were procured from the American Type Cell Culture (ATCC) in Manassas (MAX), VA, USA.

Techniques: MTT Assay

Journal:

Article Title: A unique ISRE, in the TATA-less human Isg20 promoter, confers IRF-1-mediated responsiveness to both interferon type I and type II

doi:

Figure Lengend Snippet: The Isg20 E-box element binds USF-1 transcription factor. Gel mobility shift assays were performed with an oligonucleotide probe containing the Isg20 E-Box element and nuclear extracts from untreated (ct) or treated Daudi cells for 2 h with either IFN type I (I) or IFN type II (II). The antibodies against USF-1 or c-Myc used for the supershift are indicated at the top of the gel. The USF-1 complex and supershifted complex is indicated. The E-box probe used is indicated beneath the gel.

Article Snippet: Polyclonal rabbit antisera against Stat1α p91 (sc-345), Stat2, Stat3, IRF-1 (sc-497), USF-1 (sc-229) and monoclonal mouse antibody against c-Myc (sc-42) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Mobility Shift

Journal: Cancers

Article Title: Targeting p53 for Melanoma Treatment: Counteracting Tumour Proliferation, Dissemination and Therapeutic Resistance

doi: 10.3390/cancers13071648

Figure Lengend Snippet: SLMP53-2 inhibits melanoma cell growth through induction of cell cycle arrest and apoptosis. ( A ) IC 50 values of SLMP53-2 in A375, G361, MEWO and SK-MEL-5 melanoma cells obtained by colony formation assay for 11 days; data were normalized to DMSO and correspond to mean ± SEM, n = 5 (two replicates each). ( B ) Colony formation assay for A375, G361, MEWO and SK-MEL-5 melanoma cells treated with SLMP53-2 for the indicated concentrations. Images are representative of five independent experiments. ( C ) Effect of SLMP53-2 on growth and morphology of A375 cells for the indicated time points; images are representative of five independent experiments (scale bar = 100 μm, magnification = ×100). ( D ) Apoptosis (Annexin V-positive cells) was evaluated in A375 cells after 24, 48 and 72 h of treatment with 12 μM SLMP53-2. ( E ) Cell cycle analysis in A375 cells was determined after 24, 48 and 72 h of treatment with 12 μM SLMP53-2. In ( D , E ), data are mean ± SEM, n = 5; values are significantly different from DMSO: * p < 0.05, one-way ANOVA followed by Tukey’s test. ( F , G ) Effect of SLMP53-2 on three-day-old A375 spheroids, for up to 8 days of treatment. In G , data are mean ± SEM, n = 5; values are significantly different from DMSO: * p < 0.05, one-way ANOVA followed by Tukey’s test. ( H , I ) Evaluation of spheroid formation after 10 days of treatment with SLMP53-2; treatment was performed at the seeding time of A375 cells. In I , data are mean ± SEM, n = 5; values are significantly different from DMSO: * p < 0.05, one-way ANOVA followed by Tukey’s test. In ( F , H ), images are representative of five independent experiments; scale bar = 100 μm; magnification = 100×.

Article Snippet: Human melanoma A375 (CLS Cat# 300110/p852_A-375, RRID:CVCL_0132) and SK-MEL-5 (CLS Cat# 300157/p634_SK-MEL-5, RRID:CVCL_0527) cells were purchased from CLS Cell lines service (Eppelheim, Germany).

Techniques: Colony Assay, Cell Cycle Assay

Journal: Cancers

Article Title: Targeting p53 for Melanoma Treatment: Counteracting Tumour Proliferation, Dissemination and Therapeutic Resistance

doi: 10.3390/cancers13071648

Figure Lengend Snippet: SLMP53-2 inhibits melanoma cell migration and invasion. ( A ) A375 and SK-MEL-5 confluent cells were treated with 2 or 4 μM SLMP53-2, respectively; cells were observed at 24 and 32 h (A375) and 30 and 48 h (SK-MEL-5) in the wound-healing assay. Images are representative of five independent experiments; scale bar = 100 μM; magnification = 100×. ( B ) Quantification of wound closure using randomly selected microscopic fields (six fields per sample). Data are mean ± SEM, n = 5; values are significantly different from DMSO: * p < 0.05, two-way ANOVA followed by Sidak’s test. ( C ) Effect of 2 μM SLMP53-2 on migration of A375 and SK-MEL-5 cells after 24 h of treatment. The relative number of migratory cells was determined by analysis of fluorescence signal intensity; values with DMSO were set as 1. Data are mean ± SEM, n = 5 (two replicates each); values are significantly different from DMSO: * p < 0.05, Student’s t -test. ( D ) Effect of 2 μM SLMP53-2 on the invasion of A375 and SK-MEL-5 cells after 24 h of treatment. Cells able to invade through an ECMatrix layer were quantified by fluorescence signal; values with DMSO were set as 1. Data are mean ± SEM, n = 5 (two replicates each); values are significantly different from DMSO: * p < 0.05, Student’s t -test. ( E ) Effect of SLMP53-2 on lactate secretion by A375 and SK-MEL-5 cells after 8 h of treatment. Cell density for each sample was used to normalize relative luminescence units (RLU) signal. Data are mean ± SEM, n = 5 (two replicates each); values are significantly different from DMSO: * p < 0.05; unpaired Student’s t -test.

Article Snippet: Human melanoma A375 (CLS Cat# 300110/p852_A-375, RRID:CVCL_0132) and SK-MEL-5 (CLS Cat# 300157/p634_SK-MEL-5, RRID:CVCL_0527) cells were purchased from CLS Cell lines service (Eppelheim, Germany).

Techniques: Migration, Wound Healing Assay, Fluorescence

Journal: Cancers

Article Title: Targeting p53 for Melanoma Treatment: Counteracting Tumour Proliferation, Dissemination and Therapeutic Resistance

doi: 10.3390/cancers13071648

Figure Lengend Snippet: SLMP53-2 interferes with key molecular players in epithelial-to-mesenchymal transition (EMT) and angiogenesis. ( A – D ) Protein expression levels of crucial regulators of EMT and angiogenesis in A375 ( A , B ) and SK-MEL-5 ( C , D ) melanoma cells after 48 h of treatment with SLMP53-2 (in A375 cells, β-catenin was detected for 8 h and E-cadherin and TWIST for 24 h of treatment). Immunoblots are representative of five independent experiments; GAPDH was used as a loading control. In ( B , D ), quantification of protein expression levels is shown; values with DMSO were set as 1; data are means ± SEM, n = 5.

Article Snippet: Human melanoma A375 (CLS Cat# 300110/p852_A-375, RRID:CVCL_0132) and SK-MEL-5 (CLS Cat# 300157/p634_SK-MEL-5, RRID:CVCL_0527) cells were purchased from CLS Cell lines service (Eppelheim, Germany).

Techniques: Expressing, Western Blot, Control