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. 2020 Apr 30;295(24):8331–8347. doi: 10.1074/jbc.RA120.012355

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© 2020 Zoltner et al.

Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license.

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Figure 7. — Suramin affects levels of cytosolic and mitochondrial ATP. A and B, subcellular localization of luciferase_v5 (A) and MLS_luciferase_v5 (with N-terminal mitochondrial localization signal) (B) was determined in BSF whole-cell lysates (WCL) and the corresponding organellar (ORG) and cytosolic (CYT) fraction separated by digitonin extraction. Purified fractions were analyzed by Western blotting with the following antibodies: anti-luciferase, anti-mt Hsp70 (mitochondrial marker), and anti-adenosine phosphoribosyltransferase (APRT) (cytosolic marker). The relevant sizes of the protein marker are indicated on the left. C and D, the cytosolic and mitochondrial ATP content was measured by a TECAN M200 plate reader upon the addition of luciferin to living cells. The y axis represents the percentage of the measured light units of cells treated with suramin to BSF cells, relative to those that were not treated with suramin (0-h treatment). The displayed results represent the average maximal signals obtained from three independent measurements. Error bars, S.E.; ****, p < 0.001.