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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, <t>Z-VAD-FMK;</t> HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.
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Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, Z-VAD-FMK; HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.

Journal: Aging and Disease

Article Title: Lanatoside C, a Novel Senolytic, Ameliorates Atherosclerosis in Mice

doi: 10.14336/AD.2025.1219

Figure Lengend Snippet: Senolytic mechanism of Lana C in RS HUVECs. Young or RS HUVECs were pretreated with fluorescent probe and KCl, and then Lana C treated ( A-C ). (A) DiBAC4(3) fluorescent microscopy. (B) DiBAC4(3) fluorescence intensity for plasma membrane potential in young or RS HUVECs (n=3 in each group). (C) Cal-520® AM fluorescence intensity for intracellular Ca 2+ in young or RS HUVECs (n=3 or 8 in each group). RS HUVECs were pretreated with KCl (10 mM), amiloride (100 μM), ZVF (20 μM), and ABT-263 (100 nM) and then treated with Lana C ( D-G ). (D-E) Cell viability measured by cell counting (n=3 or 4 in each group). (F) Western blotting of active caspase-3 protein. G. Quantification of the levels of active caspase-3 protein (n=3 in each group). (H) Schematic illustration of Lana C’s mechanism of action. Values are presented as means ± SEM and data were analyzed with one-way ANOVA followed by a post-hoc test. * p <0.05, ** p <0.01. Abbreviations: Y, young; RS, replicatively senescent; ABT, ABT-263; ZVF, Z-VAD-FMK; HUVEC, human umbilical vein endothelial cell; DiBAC4(3), bis-(1,3-dibutylbarbituric acid) trimethine oxonol; KCl, potassium chloride.

Article Snippet: Z-VAD-FMK was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Microscopy, Fluorescence, Clinical Proteomics, Membrane, Cell Counting, Western Blot