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. 2018 May 4;9:160. doi: 10.3389/fgene.2018.00160

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Copyright © 2018 Warnsmann, Hainbuch and Osiewacz.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Isorhamnetin do not affect respiration and ROS metabolism of the P. anserina WT. (A) Complex I-dependent OCR of mitochondria from WT cultures treated with isorhamnetin (Isor, n = 4 biological replicates, each with three to six technical replicates) compared to mitochondria from DMSO-treated cultures (Con; n = 3 biological replicates, each with five technical replicates). State 4 OCR of DMSO-treated mitochondria was set to 100%. (B) Representative BN-PAGE analysis of isolated mitochondria from WT cultures treated with isorhamnetin (Isor) compared to DMSO (Con) treated. The CI₁CIII₂CIV_0-2 (S_0-2) supercomplexes, dimeric complexes III and V (III₂ and V₂) as well as monomeric complexes I₁, IV₁, and V₁ are visualized by Coomassie staining. (C) Densitometric quantification of relative protein levels of ETC complexes and supercomplexes in mitochondrial extracts from isorhamnetin-treated cultures compared to DMSO-treated cultures. Optical densities of the different complexes were normalized to the total Coomassie staining of the corresponding lane. The mean abundance in the DMSO control was set to 1. (D) Qualitative determination of superoxide anion and hydrogen peroxide release in P. anserina WT cultures (n = 4) treated with isorhamnetin (Isor) or DMSO (Con) by a histochemical staining protocol using NBT and DAB. (E) Quantification of H₂O₂ release measurement from P. anserina PaMth1_OEx cultures treated with quercetin (Quer; n = 4) compared to DMSO (Con; n = 4) treatment. The mean release of the DMSO-treated cultures was set to 1. (F) Representative “in-gel” SOD (PaSOD1 and PaSOD2) activity staining, (G) representative “in-gel” peroxidase activity staining from total protein extract of P. anserina WT cultures treated with isorhamnetin (Isor) or DMSO (Con).