Extended Data Fig. 5: Catalytic activity of MOF regulates levels of LCFAs.
a, Quantification of mitochondrial DNA via genomic qPCR. n = 4 brains per genotype. b, Mitochondrial respiration in cells acutely isolated from E14.5 brains and analysed using the Seahorse analyser. n = 3 Mof-nKO, 2 Moffl/+ Nes-CreT/+, 8 Cre-negative. A–antimycin A, F–FCCP, O–oligomycin, R–rotenone. c, Quantification of basal respiration rate from Seahorse profiles (panel b). d, Quantification of ATP-linked respiration from Seahorse profiles (panel b). e, Mof mRNA levels in Mof-KO and WT neurosphere cultures following 4-hydroxy-tamoxifen (4OHT) treatment. Exp.1 n = 6 Mof-KO, 8 WT; Exp.2 n = 5 Mof-KO, 8 WT neurosphere cultures. f, Western blot analysis of MOF, Flag-tagged MOF, Actin, H4K16ac and histone H3 protein levels after re-expression of WT and E350Q MOF in Mof-KO and control MEFs. L indicates ladder. Protein sizes are indicated in kDa. 4 independent MEF lines per genotype and per treatment were analysed. g-k, Quantification of g, Flag expression, h Total MOF, i endogenous MOF, j exogenous MOF and k H4K16ac levels in WT and Mof-KO cultures. Quantification was carried out from the blots shown in panel f. Total MOF levels are a combination of Flag-tagged MOF (exogenous, higher MW) and endogenous MOF. Flag-tagged and endogenous MOF levels were standardized to Actin. H4K16ac levels were standardized to histone H3. Asterisks signify statistical significance at *p < 0.05, **p < 0.01 and ***p < 0.001. Precise p-values are provided in Source Data Extended Data Fig. 5. n = 4 independent MEF lines per genotype and per treatment. l LCFA levels in WT and Mof-KO MEFS after the re-expression of WT and E350Q mutant MOF. 4 MEF lines per genotype and per treatment were analysed. Data are presented as mean ± s.e.m. and were analysed using a two-tailed Student’s t-test. Statistical source data and unprocessed blots are shown in Source Data Extended Data Fig. 5.
