Figure 9 | Scientific Reports

Figure 9

From: Loss of the Mitochondrial Fatty Acid β-Oxidation Protein Medium-Chain Acyl-Coenzyme A Dehydrogenase Disrupts Oxidative Phosphorylation Protein Complex Stability and Function

Figure 9

Identification of MCAD interacting proteins. (A) Co-immunoprecipitation (Co-IP) and mass-spectrometry analysis of 143B cell mitochondria. Crude mitochondria were solubilised in 1% (v/v) Triton X-100 and proteins incubated with Protein A-sepharose coupled MCAD antibodies or protein A-sepharose alone. Eluates were analyzed by label-free quantitative mass-spectrometry (LFQ). Log2 LFQ intensities were submitted to a modified two-sided two-sample t-test with significance determined through permutation-based false discovery rate (FDR) statistics. Closed circles, mitochondrial protein; empty circles, non-mitochondrial protein; black, proteins with <1% FDR. (B) As for (A) using either control 143B cell mitochondria or mitochondria from 143B MCAD knockout (KO) cells solubilised in 1% (v/v) Triton X-100, bound to Protein A-sepharose coupled MCAD antibodies in both cases. (C) As for (B) using either control 143B mitochondria or mitochondria from 143B MCAD knockout (KO) cells solubilised in 1% (w/v) digitonin, bound to Protein A-sepharose coupled MCAD antibodies in both cases. (D) As for (A) using HepG2 control cell mitochondria and 1% (w/v) digitonin, bound to Protein A-sepharose coupled MCAD antibodies or protein A-sepharose alone.

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