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RA413S induces mitochondrial damage and alters cellular bioenergetics.

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posted on 10.09.2021, 17:36 by Ravi K. Anchoori, Logan George, Ssu-Hsueh Tseng, Brandon Lam, Srinidhi Polkampally, Anjali D. Amiano, Palmer Foran, Hannah Tsingine, Harideep Samanapally, Fernanda Carrizo Velasquez, Samarjit Das, Deyin Xing, Ahmad Bin Salam, Balasubramanyam Karanam, Chien-Fu Hung, Richard B. S. Roden

Fluorescent microscopy images of (A) HeLa (B) BR5 cells treated with RA413S for 18 h at the indicated concentrations. Cells were fixed and stained with MitoTracker (red [A]/yellow [B] pseudocoloring and Hoechst (blue). Fragmented mitochondria is visible in RA413S treated samples (C) Assessment of the impact of treatment upon intracellular ATP levels using bioluminescence of ES2 cells expressing firefly luciferase after addition of D-luciferin; RA183 and RA413S treatment caused reduction ATP-dependent luminescence. (D-I) Seahorse assay in ES2 cells upon treatment with RA413S (100 nM) and RA414 (100 nM) or vehicle (DMSO) for 12 h. (D-J) Oxygen consumption and extracellular acidification rates by ES2 cells upon treatment with 500 nM of RA413S and 100 nM RA414 or vehicle (DMSO) for 12 hr. Mitochondrial function was measured following sequential additions of oligomycin, FCCP, Rotenone and Antimycin A into the culture media. (E-J) Oxygen consumption rate (OCR) in ES2 cells as measured by a Seahorse XF Analyzer. Data are representative of 3 independent experiments, and 9 biologic replicates per conditions were used.

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