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. 2018 Feb 15;8:3081. doi: 10.1038/s41598-018-21361-6

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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SDHB knockdown causes severe defects in mitochondrial respiration but increases glucose uptake and glycolytic metabolism. (A) ATP assay in SDHB-knockdown cells. Total extracts prepared from cells as indicated were subjected to an ATP assay using the ATP Bioluminescence Assay Kit CLSII. (B,C) Δψ_m assay of SDHB-knockdown cells. Cells as indicated were stained with Rhodamine123 and TMRM and then analyzed using a flow cytometer. (D,E) ROS assay of SDHB-knockdown cells. Cells were treated with CM-H₂DCFDA and DCF as indicated and subsequently analyzed using a flow cytometer. (F) Glucose uptake assay of SDHB-knockdown cells. Cells as indicated were loaded with 2-NBDG and then analyzed using a flow cytometer. (G) LDH activity assay of SDHB-knockdown cells. Total extracts prepared from cells were subjected to a LDH activity assay using the CytoTox 96^® Non-Radioactive Cytotoxicity Assay. (H) Color change and pH measurements of the conditioned media cultured with SDHB-knockdown cells. Cells as indicated were cultured until confluent and then incubated in fresh media for 12 h. The color and pH of the conditioned media were imaged and measured. The *^, ** and *** represented P-value < 0.05, <0.01 and <0.005, respectively.