Fig. 3: Downregulation of ATP6V1C2 expression inhibits the induction of ferroptosis in normal cells. | Nature Communications

Fig. 3: Downregulation of ATP6V1C2 expression inhibits the induction of ferroptosis in normal cells.

From: Senescence-associated lysosomal dysfunction impairs cystine deprivation-induced lipid peroxidation and ferroptosis

Fig. 3

a Lysosomal pH in presenescent (Control) and senescent TIG-3 (Irrad and Rep) cells measured by LysoSensor. b–f Presenescent TIG-3 cells were treated with 1 µM concanamycin C (ConC) for 24 h and subjected to lysosomal pH analysis (b) or lipid peroxidation measurement via C11-BODIPY fluorescence (c). The ratio of oxidized to reduced BODIPY (BODIPYox/BODIPYred) calculated from the median fluorescence intensity is shown. The data represent the mean ± s.d. of three independent experiments. d, e Cell viability of and LDH release by TIG-3 cells after 24 h of treatment with 5 µM erastin with or without 1 µM ConC. f Percentage of peroxidized PG, Data are presented as the mean ± s.d. of n  =  3 biological replicates. g Heatmap showing changes in the gene expression of V-ATPase components in presenescent (Control) and senescent TIG-3 (Irrad and Rep) cells. h Schematic of V-ATPase. Created in BioRender. Takahashi, A. (2025) https://BioRender.com/lhkt32a. Presenescent TIG-3 cells were subjected to transfection with the indicated siRNA oligos twice (at 2-day intervals). These cells were then subjected to RT‒qPCR analysis (i) and lysosomal pH was measured via LysoSensor (j). Cell viability (k) of and LDH release (l) by siControl- or siATP6V1C2-transfected TIG-3 cells after 24 h of treatment with 5 µM erastin. Data are presented as the mean ± s.d. of n  =  3 biological replicates (c–e, k, l). For (a, b, j) the box represents the interquartile range (IQR), with a horizontal line and a black point indicating the median and mean, respectively. Whiskers extend to the most extreme data points within 1.5 times the IQR from the quartiles. White points beyond this range represent outliers. Statistical analysis was performed via the Kruskal-Wallis test followed by Dunn’s test (a, j), the two-sided Mann-Whitney U test (b), and one-way ANOVA with Tukey’s test (c, d, e, f, k, l), and one-way ANOVA with Dunnett’s test (i). Source data are provided as a Source Data file.

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