Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: Mitochondrial function in HCE-2 cells following exposure to 100 µg/mL PM 10 with or without 50 nM SKQ1 was evaluated using a Seahorse bioanalyzer after 24 h. A representative profile of the oxygen consumption rate (OCR) shows a reduced OCR after PM 10 exposure compared to the control and SKQ1 groups, which was improved by SKQ1 ( A ). PM 10 significantly reduced basal respiration ( B ), maximum respiration ( C ), spare capacity ( D ), ATP production ( E ), and coupling efficiency ( F ). SKQ1 pre-treatment significantly reversed these effects. Significant changes are denoted by p -values indicated by symbols: * p < 0.05; ** p < 0.01; *** p < 0.001. n = 3 biological replicates for each group.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Control

Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: JC-1 staining in HCE-2 cells following exposure to 100 µg/mL PM 10 in the absence or presence of 50 nM SKQ1. Green fluorescence (monomers) reflects a reduced mitochondrial membrane potential, whereas red fluorescence indicates healthy mitochondria. PM 10 -exposed cells had a lower mitochondrial membrane potential than the control or SKQ1 alone after 24 h. Pre-treatment with SKQ1 before PM 10 exposure partially restored the mitochondrial membrane potential. CCCP was used as a positive control. Arrows indicate depolarized mitochondria. n = 3 biological replicates were used for each group in this study.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Staining, Fluorescence, Membrane, Control, Positive Control

Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: The mitochondrial membrane potential in HCE-2 cells was assessed by flow cytometry using JC-1 dye following exposure to 100 µg/mL PM 10 with or without 50 nM SKQ1. Scatter plots ( A ) and graphical quantification ( B ) show that PM 10 -exposed cells have more depolarized mitochondria (lower mitochondrial membrane potential) than the control or SKQ1 alone (normal mitochondria) after 24 h. Pre-treatment with SKQ1 before PM 10 exposure significantly reversed these changes. CCCP was used as a positive control. *** p < 0.001; n = 3 biological replicates were used for each group in this study.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Membrane, Flow Cytometry, Control, Positive Control

Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: qRT-PCR in HCE-2 cells exposed to 100 µg/mL PM 10 in the presence or absence of 50 nM SKQ1. Compared to the control and SKQ1 groups, PM 10 exposure significantly downregulated MFN1 ( A ), MFN2 ( B ), OPA1 ( C ), PGC-1α ( F ), TFAM ( G ), and COX4i1 ( H ) but upregulated FIS1 ( D ) and DRP1 ( E ) after 24 h. SKQ1 pre-treatment reversed these effects. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 3 biological replicates were used for each group in this study.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Quantitative RT-PCR, Control

Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: Protein analysis in HCE-2 cells exposed to 100 µg/mL PM 10 in the presence or absence of 50 nM SKQ1 using the Protein Simple Abby system. PM 10 exposure significantly downregulated MFN1 ( A ), TFAM ( B ), COX4i1 ( C ), and PGC-1α ( D ) compared to the control and SKQ1 alone after 24 h. Pre-treatment with SKQ1 before PM 10 exposure reversed these effects. * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3 biological replicates were used for each group in this study.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Control

Journal: Antioxidants

Article Title: PM 10 Disrupts Mitochondrial Homeostasis in Corneal Epithelial Cells: Protective Effects of SKQ1

doi: 10.3390/antiox15030284

Figure Lengend Snippet: Representative images of mitochondrial morphology ( A ) and DNA copy number ( B ) after exposure of HCE-2 cells to 100 µg/mL PM 10 ± 50 nM SKQ1. PM 10 exposure induced mitochondrial fragmentation and reduced mtDNA copy number compared to control and SKQ1 alone after 24 h. SKQ1 pre-treatment partially restored normal elongated morphology and increased reduction in DNA copy number. n = 3 biological replicates were used for each group in this study. The white frames indicate enlarged areas to clearly illustrate the differences in morphology. * p < 0.05, *** p < 0.001.

Article Snippet: HCE-2 cells were grown on 12 mm coverslips coated FNC (, Athena ES, Baltimore, MD, USA) in a 12-well plate for 24 h; pre-treated with SKQ1; and exposed to PM 10 , as described in the Tissue Culture and Treatments Section.

Techniques: Control

Journal: Drug Metabolism and Disposition

Article Title: In Vitro Metabolism and Drug-Drug Interaction Potential of UTL-5g, a Novel Chemo- and Radioprotective Agent

doi: 10.1124/dmd.114.060095

Figure Lengend Snippet: Kinetic parameters for UTL-5g hydrolysis to DCA and ISOX in HLM and by hCE1b and hCE2

Article Snippet: BD UltraPool HLM 150 (150-donor human liver microsome pools with equal gender mix), recombinant hCE1b and hCE2, CYP1A2 Supersomes, as well as NADPH regenerating system solution A and B were obtained from Corning Inc. (Woburn, MA).

Techniques:

Journal: Drug Metabolism and Disposition

Article Title: In Vitro Metabolism and Drug-Drug Interaction Potential of UTL-5g, a Novel Chemo- and Radioprotective Agent

doi: 10.1124/dmd.114.060095

Figure Lengend Snippet: UTL-5g hydrolysis to ISOX and DCA in hCE1b (A) and hCE2 (B) in the absence of NADPH. Each data point represents a single measurement. The curve represents the fitting of both ISOX and DCA formation rate versus initial UTL-5g concentration pooled data from two independent experiments to the Michaelis-Menten equation.

Article Snippet: BD UltraPool HLM 150 (150-donor human liver microsome pools with equal gender mix), recombinant hCE1b and hCE2, CYP1A2 Supersomes, as well as NADPH regenerating system solution A and B were obtained from Corning Inc. (Woburn, MA).

Techniques: Concentration Assay