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. 2023 Jun 10;42:145. doi: 10.1186/s13046-023-02718-w

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — MiR-494 affects redox homeostasis in HCC cells. A ROS production in control (pMXs) and miR-494-overexpressing Huh-7 cells measured using dichlorofluorescin diacetate (DCFDA) and (B) mitochondrial superoxide production measured using the mitochondrial specific probe MitoSOX Red. Y-axes reports the arbitrary fluorescence units normalized to controls. Mean ± SD values are displayed. Three independent experiments were performed. C Representative confocal microscope images of control (pMXs) and miR-494-overexpressing Huh-7 cells stained with Fluo-3 AM. Violin plot reports the quantification of Fluo-3 intensity emission per cell normalized to control. Five randomly selected fields were analysed in two independent experiments. Mean ± SD values are displayed. Scale bar = 30 µm. D Lipid membrane peroxidation status in control (pMXs) and miR-494-overexpressing Huh-7 cells measured using the lipid peroxidation sensor BODIPY™ 581/591 C11. The oxidation of the dye results in a green-shift fluorescence emission peak. Y-axis reports quantification of the Red/Green intensity ratio per cell normalized to control. Five randomly selected fields were analysed in three independent experiments. Mean ± SD values are displayed. Scale bar, 30 µM. E Glutathione content in control (pMXs) and miR-494-overexpressing Huh-7 cells. Y-axis reports the luminescence signal relative to glutathione levels normalized to control. Mean ± SD values are displayed. Three independent experiments were performed. F ROS production in control (pMXs) and miR-494-overexpressing cells after 24 h of exposure to no-glucose medium measured using DCFDA. Data are presented as fold change of ROS production in no-glucose with respect to normal culture condition (RPMI). Three independent experiments were performed. G Real time PCR analysis of PGC-1alpha and (H) HAO2 mRNA levels in control (pMXs) and miR-494-overexpressing Huh-7 cells. Y-axis reports 2^−ΔΔ.^Ct values corresponding to mRNA levels. Normalized mean ± SD values are displayed. Beta-actin was used as housekeeping gene. Real Time PCR analysis was performed in two independent experiments in triplicate. The statistical analysis was performed using two-tailed unpaired Student's t-test. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001