Journal: Antioxidants

Article Title: Conjugated Linolenic Acids Induce Ferroptosis in Human and Zebrafish Melanoma Cells

doi: 10.3390/antiox15030360

Figure Lengend Snippet: Effects of ACSL4 or GPX4 inhibitors on the PunA-induced cytotoxicity in melanoma cells. Viability of A375, WM266.4 and ZMEL1 cells treated with punicic acid (PunA) was assessed in the presence of increasing doses of PRGL493 ( a – c ), RSL3 ( d – f ) or ML210 ( g – i ). Relative cell viability was normalized to control cells that were cultured with medium without any added inhibitor or CLnA, defined as 100%. For the cells cultured without CLnA, the culture medium still included the vehicles used for PRGL493, RSL3 and ML210. Data are presented as mean ± standard error of the mean (SEM) of 3 independent experiments. Dose-response curves have been fitted to the data. Statistical significance was assessed by two-way ANOVA with Dunnett’s test, comparing different tested inhibitor concentrations (0.03, 0.1, 0.3, 1, 3, 10 μM) against 0 μM. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****). Only significant differences are shown.

Article Snippet: Additional 96-well plates were subjected to either 2.5 μM PunA (A375 and WM266.4 cells) or 10 μM PunA (ZMEL1 cells), in combination with an increasing dose of the ACSL4 inhibitor PRGL493 (HY-139180, MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Control, Cell Culture

Journal: Antioxidants

Article Title: Conjugated Linolenic Acids Induce Ferroptosis in Human and Zebrafish Melanoma Cells

doi: 10.3390/antiox15030360

Figure Lengend Snippet: Modulation of acsl4a and gpx4b gene expression in ZMEL1 cells treated with fatty acids. The fold changes in relative quantity (RQ) were calculated for acsl4 a ( a ) or gpx4 b ( b ) in cells exposed to 20 μM of a tested fatty acid for 24 h, relative to the control cells, which were cultured with DMEM without any added fatty acid. The following fatty acids were tested: α-linolenic acid (ALA), oleic acid (OLA), punicic acid (PunA), α-eleostearic acid (α-ESA), β-eleostearic acid (β-ESA) and jacaric acid (JA). The RQ was calculated as RQ = 2 −ΔCt , the ΔC t value representing the difference in C t value between the target gene and a panel of reference genes, which are actin beta 2, beta-2-microglobulin, hypoxanthine phosphoribosyl transferase 1 and TATA-binding protein. Data are presented as mean ± standard error of the mean (SEM) of 3 independent experiments (N = 3, n = 3). Significance was assessed by one-way ANOVA with Tukey’s multiple comparison test among treatments. Significant differences between the fatty acid-treated groups and the control are marked above the columns. p < 0.05 (*); p < 0.01 (**); p < 0.001 (***), p < 0.0001 (****). Only significant differences are shown.

Article Snippet: Additional 96-well plates were subjected to either 2.5 μM PunA (A375 and WM266.4 cells) or 10 μM PunA (ZMEL1 cells), in combination with an increasing dose of the ACSL4 inhibitor PRGL493 (HY-139180, MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Gene Expression, Control, Cell Culture, Binding Assay, Comparison