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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence <t>microscope</t> (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.
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Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence microscope (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.

Journal: Journal of Lipid Research

Article Title: The fate of intracellular S1P regulates lipid droplet turnover and lipotoxicity in pancreatic beta-cells

doi: 10.1016/j.jlr.2024.100587

Figure Lengend Snippet: Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence microscope (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus untreated, # P < 0.05, ## P < 0.01, versus INS1E-ctr cells treated in the same way.

Article Snippet: Cells were cultured for 48 h and afterwards exposed to FFAs for 24 h. Live cell imaging was performed using a CFP-YFP dual filter (excitation, 427 nm and 504 nm; emission, 520 nm) with a Olympus IX 81 inverted microscope system and the CellSens software (Olympus, Hamburg, Germany) for imaging and analysis.

Techniques: Incubation, Expressing, Fluorescence, Software, Microscopy

Effects of enhanced S1P turnover and FFA on peroxisomal and mitochondrial hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. Peroxisomal hydrogen peroxide generation was estimated by expressing the fluorescence sensor HyPer-Peroxi (A) and mitochondrial by expressing HyPer-Mito (B) sensor proteins and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence microscope (representative pictures from n = 3 individual experiments). Arrows depict representative cells with higher red HyPer protein fluorescence. Bars: 10 μm.

Journal: Journal of Lipid Research

Article Title: The fate of intracellular S1P regulates lipid droplet turnover and lipotoxicity in pancreatic beta-cells

doi: 10.1016/j.jlr.2024.100587

Figure Lengend Snippet: Effects of enhanced S1P turnover and FFA on peroxisomal and mitochondrial hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. Peroxisomal hydrogen peroxide generation was estimated by expressing the fluorescence sensor HyPer-Peroxi (A) and mitochondrial by expressing HyPer-Mito (B) sensor proteins and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence microscope (representative pictures from n = 3 individual experiments). Arrows depict representative cells with higher red HyPer protein fluorescence. Bars: 10 μm.

Article Snippet: Cells were cultured for 48 h and afterwards exposed to FFAs for 24 h. Live cell imaging was performed using a CFP-YFP dual filter (excitation, 427 nm and 504 nm; emission, 520 nm) with a Olympus IX 81 inverted microscope system and the CellSens software (Olympus, Hamburg, Germany) for imaging and analysis.

Techniques: Incubation, Expressing, Fluorescence, Software, Microscopy