Fig. 5: Vitamin A eliminates lipid peroxidation and upregulates major anti-ferroptotic regulators in HT-1080 as well as in cortical neurons. | Nature Communications

Fig. 5: Vitamin A eliminates lipid peroxidation and upregulates major anti-ferroptotic regulators in HT-1080 as well as in cortical neurons.

From: Suppression of ferroptosis by vitamin A or radical-trapping antioxidants is essential for neuronal development

Fig. 5

a Flow cytometry of C11-BODIPY staining: (left) histograms and (right) intensity of HT-1080 cells co-treated with the ferroptosis inducer (RSL3), and inhibitor (Fer-1) or vitamin A (ATRA). Data are % intensity of median fluorescence normalized to RSL3-treated cells ± SD of n = 4 biologically independent replicates; one-way-ANOVA with Tukey’s test. b Immunostaining of 4-Hydroxynonenal (4-HNE) and flow cytometry: (left) histograms and (right) intensity of HT-1080 cells co-treated with the ferroptosis inducer (RSL3), and inhibitor (Fer-1) or vitamin A (ATRA). Data are % intensity of median fluorescence normalized to RSL3-treated cells ± SD of n = 3 biologically independent replicates; one-way-ANOVA with Tukey’s test. c TBARS assay of HT-1080 cells co-treated with the ferroptosis inducer (RSL3) and inhibitor (Fer-1) or vitamin A (ATRA). Data are mean ± SD of n = 4 biologically independent replicates; one-way-ANOVA with Tukey’s test. d mRNA expression, using quantitative RT-PCR, of various anti-ferroptotic regulators in HT-1080 cells treated with DMSO or 10 µM vitamin A (ATRA) and 10 µM RARi. Data are mean ± SD of n = 3 biologically independent replicates; one-way-ANOVA with Tukey’s test. e protein expression, using Western Blot, of various anti-ferroptotic regulators (GPX4, FSP1, ACSL3, SCD1) as well as pro-ferroptotic regulators (ACSL4, LPCAT3) in HT-1080 cells treated with DMSO or 20 µM vitamin A (ATRA). n = 3 biologically independent replicates are shown. f mRNA expression (qRT-PCR) of various anti-ferroptotic regulators in cortical neurons differentiated with low or high vitamin A. Data plotted are mean ± SD of n = 3 biologically independent replicates; unpaired t test, two-tailed. g protein expression, using Western Blot, of various anti-ferroptotic regulators (GPX4, FSP1, SCD1) in immature neurons differentiated with low or high vitamin A. n = 3 biologically independent replicates are shown. Source data are provided as a Source Data file.

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