sj-pdf-1-jcb-10.1177_0271678X221077341 - Supplemental material for Characterizing region-specific glucose metabolic profile of the rodent brain using Seahorse XFe96 analyzer

Supplemental material, sj-pdf-1-jcb-10.1177_0271678X221077341 for Characterizing region-specific glucose metabolic profile of the rodent brain using Seahorse XFe96 analyzer by Linshu Wang, Kiran Chaudhari, Ali Winters, Yuanhong Sun, Ran Liu and Shao-Hua Yang in Journal of Cerebral Blood Flow & Metabolism</p

Effects of E2, PPT, DPN, ZYC-26 and ZYC-23 on intracellular lipid peroxidation in BSO-treated FRDA fibroblasts.

<p>All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells. 1.0 normalized 8-isoprostane control concentration = 6.23 pg/mL.</p

Effects of E2, PPT, DPN, ZYC-26 and ZYC-23 on the activity of aconitase in BSO-treated FRDA fibroblasts.

<p>All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells.</p

Effects of E2 and ZYC-26 on mitochondrial function in BSO-treated FRDA fibroblasts.

<p>A.) Oxygen consumption rate (OCR; in pMoles/min) B.) Basal respiratory rate (in pMoles/min) C.) Maximal respiratory rate (in pMoles/min) All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells.</p

Proposed mechanism of 17β-Estradiol in BSO-treated FRDA fibroblasts.

<p>Proposed mechanism of 17β-Estradiol in BSO-treated FRDA fibroblasts.</p

Effects of E2, PPT, DPN, ZYC-26 and ZYC-23 on the collapse of mitochondrial membrane in BSO-treated FRDA fibroblasts.

<p>All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells.</p

Figure 3

<p>A.) Calcein AM imaging demonstrating cell viability between vehicle control and BSO treatment groups at 24, 36 and 48 hours. Scale bar = 200 µm. B.) Effects of E2, PPT, DPN, ZYC-26 and ZYC-23 on cell viability in BSO-treated FRDA fibroblasts. All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells.</p

Structures of compounds assessed for protection against BSO toxicity in FRDA fibroblasts.

<p>Structures of compounds assessed for protection against BSO toxicity in FRDA fibroblasts.</p

Effects of E2, PPT, DPN, ZYC-26 and ZYC-23 on the intracellular ATP concentration inside of BSO-treated FRDA fibroblasts.

<p>All steroid concentrations were 100 nM, DMSO concentration was 0.1% and BSO concentration was 1 mM. Depicted are mean ± SD for n = 8 per group. * indicated p<0.05 versus BSO alone-treated cells. 100% normalized ATP control concentration = 501 pM.</p

Western blot showing the presence of small amounts of ERβ and the absence of ERα in FRDA fibroblasts compared with 661W photoreceptor cells.

<p>Western blot showing the presence of small amounts of ERβ and the absence of ERα in FRDA fibroblasts compared with 661W photoreceptor cells.</p