HEK-cFXN cells show a correlation among ROS levels, LIP and frataxin expression. HEK-cFXN cells show a correlation among ROS levels, LIP and frataxin expression.

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Wallerian Degeneration Is Executed by an NMN-SARM1-Dependent Late Ca2+ Influx but Only Modestly Influenced by Mitochondria Andrea Loreto, Michele Di Stefano,

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HEK-cFXN cells show a correlation among ROS levels, LIP and frataxin expression. HEK-cFXN cells show a correlation among ROS levels, LIP and frataxin expression. (A) The two graphs represent ROS levels analyzed concomitantly on HEK-cFXN maintained under the indicated conditions and normalized for the lowest value [in untreated cells, kept 8 days without tetracycline (Tet; dashed line)]. The inset in the left panel represents the expression of frataxin at the corresponding time points of Tet deprivation. Oxidative conditions were obtained by overnight treatment with ferric iron (50 μM ferric ammonium citrate) combined with acute exposure to hydrogen peroxide (15 min, 300 μM). The bars correspond to the mean (±s.e.m.), from three independent experiments (each condition in duplicate; >5000 cells per condition), of CellROX fluorescence signals. Representative fluorescence images from the time points indicated by the arrows are shown in the lower panel. Statistical significance was calculated using one-way ANOVA followed by Dunnett's post hoc test; *P<0.05, **P<0.01. (B) ROS levels, analyzed in wild-type HEK293 (HEK293 WT) were not affected by Tet (10 ng ml−1) treatment, even after hydrogen peroxide (15 min, 300 μM). (C) LIP was evaluated as the fold increase of calcein fluorescence after administration of 100 μM SIH, compared with before. Statistical significance was calculated using one-way ANOVA followed by Bonferroni post hoc test; *P<0.05, **P<0.01. Tommaso Vannocci et al. Dis. Model. Mech. 2018;11:dmm032706 © 2018. Published by The Company of Biologists Ltd