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USP45 suppresses ferroptosis of lung adenocarcinoma cells induced by erastin. (A–C) Biochemical assays measuring levels of MDA, Fe 2+ , GSH, and GSSG in USP45‐depleted A549 and erastin‐treated USP45‐overexpressing HCC827 cells. USP45 deficiency amplified lipid peroxidation and oxidative imbalance, while USP45 overexpression reversed these effects. (D) ROS detection <t>by</t> <t>DCF‐DA</t> staining showing elevated ROS in USP45‐deficient A549 cells and reduced ROS in USP45‐overexpressing HCC827 cells in the presence of erastin. (E) Western blot analysis of ferroptosis markers ACSL4, COX2, and FTH1, showing increased ACSL4 and COX2 expression, reduced FTH1 expression in USP45‐deficient A549 cells, and the opposite pattern in USP45‐overexpressing HCC827 cells. One‐way ANOVA followed by Tukey's HSD tests was used to determine statistical significance. Abbreviation: Era, erastin.
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USP45 suppresses ferroptosis of lung adenocarcinoma cells induced by erastin. (A–C) Biochemical assays measuring levels of MDA, Fe 2+ , GSH, and GSSG in USP45‐depleted A549 and erastin‐treated USP45‐overexpressing HCC827 cells. USP45 deficiency amplified lipid peroxidation and oxidative imbalance, while USP45 overexpression reversed these effects. (D) ROS detection <t>by</t> <t>DCF‐DA</t> staining showing elevated ROS in USP45‐deficient A549 cells and reduced ROS in USP45‐overexpressing HCC827 cells in the presence of erastin. (E) Western blot analysis of ferroptosis markers ACSL4, COX2, and FTH1, showing increased ACSL4 and COX2 expression, reduced FTH1 expression in USP45‐deficient A549 cells, and the opposite pattern in USP45‐overexpressing HCC827 cells. One‐way ANOVA followed by Tukey's HSD tests was used to determine statistical significance. Abbreviation: Era, erastin.
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Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe <t>H</t> <t>2</t> DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).
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Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe <t>H</t> <t>2</t> DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).
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Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe <t>H</t> <t>2</t> DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).
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Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe <t>H</t> <t>2</t> DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).
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USP45 suppresses ferroptosis of lung adenocarcinoma cells induced by erastin. (A–C) Biochemical assays measuring levels of MDA, Fe 2+ , GSH, and GSSG in USP45‐depleted A549 and erastin‐treated USP45‐overexpressing HCC827 cells. USP45 deficiency amplified lipid peroxidation and oxidative imbalance, while USP45 overexpression reversed these effects. (D) ROS detection by DCF‐DA staining showing elevated ROS in USP45‐deficient A549 cells and reduced ROS in USP45‐overexpressing HCC827 cells in the presence of erastin. (E) Western blot analysis of ferroptosis markers ACSL4, COX2, and FTH1, showing increased ACSL4 and COX2 expression, reduced FTH1 expression in USP45‐deficient A549 cells, and the opposite pattern in USP45‐overexpressing HCC827 cells. One‐way ANOVA followed by Tukey's HSD tests was used to determine statistical significance. Abbreviation: Era, erastin.

Journal: Cancer Medicine

Article Title: Ubiquitin‐Specific Protease 45 Inhibits Lung Adenocarcinoma Ferroptosis by Regulating Ubiquitination and Stability of Glutathione Peroxidase 4

doi: 10.1002/cam4.71901

Figure Lengend Snippet: USP45 suppresses ferroptosis of lung adenocarcinoma cells induced by erastin. (A–C) Biochemical assays measuring levels of MDA, Fe 2+ , GSH, and GSSG in USP45‐depleted A549 and erastin‐treated USP45‐overexpressing HCC827 cells. USP45 deficiency amplified lipid peroxidation and oxidative imbalance, while USP45 overexpression reversed these effects. (D) ROS detection by DCF‐DA staining showing elevated ROS in USP45‐deficient A549 cells and reduced ROS in USP45‐overexpressing HCC827 cells in the presence of erastin. (E) Western blot analysis of ferroptosis markers ACSL4, COX2, and FTH1, showing increased ACSL4 and COX2 expression, reduced FTH1 expression in USP45‐deficient A549 cells, and the opposite pattern in USP45‐overexpressing HCC827 cells. One‐way ANOVA followed by Tukey's HSD tests was used to determine statistical significance. Abbreviation: Era, erastin.

Article Snippet: Approximately 1.0 × 10 6 cells were treated with 10 μM DCF‐DA (50101ES01, YEASEN) in PBS for 15 min in the dark.

Techniques: Amplification, Over Expression, Staining, Western Blot, Expressing

Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe H 2 DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).

Journal: International Journal of Molecular Sciences

Article Title: Subcytotoxic Exposure to Avobenzone and Ethylhexyl Salicylate Induces microRNA Modulation and Stress-Responsive PI3K/AKT and MAPK Signaling in Differentiated SH-SY5Y Cells

doi: 10.3390/ijms27031134

Figure Lengend Snippet: Effects of exposure to AVO and EHS on cell survival and ROS formation. Cells were differentiated with retinoic acid and treated for 48 h with UV filters. Cell viability ( a , b ) was evaluated by the reduction of resazurin to resorufin. ROS formation ( c , d ) was determined using the fluorescent probe H 2 DCF-DA. Data are expressed as fold increase versus the vehicle group and reported as mean ± SD of three independent experiments (( a ): * p < 0.05 vs. 0.05; ( b ): *** p < 0.001 vs. 0.05; One-way ANOVA, post hoc Tukey test).

Article Snippet: H 2 DCF-DA, eosin, retinoic acid, leupeptin, phenylmethylsulfonyl fluoride (PMSF), β-mercaptoethanol, sodium dodecyl sulfate (SDS), dimethyl sulfoxide (DMSO), Tris·HCl and β-actin antibody were obtained from Merck Life Science S.r.L. (Darmstadt, Germany).

Techniques: