Fig. 1: TANGO2 knockout decreases mitochondrial respiration and affects mitochondrial structure and morphology in vitro.

a Immunoblots of control and TANGO2^–/– cells probed with total OXPHOS or COXII antibodies and normalised to TOMM20 (n = 6, biological replicates). b Immunoblots of proteins involved in mitochondrial translation (MRPS16, MRPL44, LRPPRC, TFAM, MRPP2) (n = 6, biological replicates). SDHA was used as a loading control. c De novo mitochondrial protein synthesis was measured by ³⁵S methionine/cystine incorporation in control and TANGO2^–/– cells. Coomassie stained gels are shown as loading controls (n = 3). d Measurement of de novo biogenesis of OXPHOS complexes by incorporation of ³⁵S-labelled cysteine and methionine. The results in c and d are representative of three independent experiments. e Oxygen consumption through the N-linked, S-linked or CIV-linked pathway using either pyruvate/glutamate/malate, succinate or TMPD/ascorbate as substrates was measured for leak (L) and OXPHOS capacity (P) in control and TANGO2^–/– cells grown in galactose media. ET capacity (ET) was measured using carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) (n = 4, biological replicates). f Membrane potential measurements in control and TANGO2^–/– cells in the presence and absence of FCCP (n = 4, biological replicates). g Mitochondrial DNA copy number in control and TANGO2^–/– cells grown in glucose was quantitated by qPCR of MT-CYB and HBB (n = 4, biological replicates). Values for (e, f and g) are means ± SD. *p < 0.05, **p < 0.01 ***p < 0.001, Student’s two-tailed t test. h Steady state levels of proteins involved in mitochondrial dynamics (LONP1, AFG3L2, CLPX, OPA1, YME1L1 and CHCHD3) (n = 6, biological replicates). TOMM20 was used as a loading control. Relative abundance of proteins shown in panels (a, b and h) was analysed relative to the loading control (n = 6). Values are means ± SD. *p < 0.05, **p < 0.01 ***p < 0.001, Student’s two-tailed t test. i Mitotracker staining of control and TANGO2^–/– cells grown in glucose and galactose media. Cells were stained with Mitotracker Orange prior to fixation and scored for fragmentation (n = 100 per cell line and treatment). Source data are provided as a Source Data file.