Fig. 6

Yy1 controls cortical metabolism and protein translation rate. a, b Oxygen consumption rate (OCR) measurement of isolated cortical cells at E12.5 using a Seahorse Extracellular Flux Analyzer reveals impaired mitochondrial bioenergetics upon ablation of Yy1. Injection of electron transport chain inhibitors are indicated after measurement 3 (oligomycin, ATP synthase inhibitor), 6 (FCCP, mitochondrial uncoupler) and 9 (Antimycin A/rotenone, complex III & I inhibitors). Parameters derived from a are indicated in b: basal respiration, ATP-linked OCR, and maximum respiration capacity. Data represented relative to first basal respiration measurement of controls and as a mean of n = 9 (control), n = 6 (Yy1cKO) error bars indicate standard error of the mean. c qRT-PCR for mitochondrial DNA content shows no difference between Yy1cKO and control cortex tissue. Graphs present mitochondrial (Mit1) versus nuclear (intergenic region, intg1) DNA ratio. d Metabolomic alterations in isolated E11.5 NPCs upon knockdown of Yy1 for 48 h. Heatmap shows enrichment of metabolic pathways which are downregulated or upregulated upon treatment with siYy1. Abbreviations: oxidative phosphorylation (OxPhos), β-oxidation (β-Ox), fatty acid (FA), metabolism (metab.), mitochondrial (mito.), degradation (degrad.). e–h Knockdown of Yy1 reduces protein translation rate. OP-puro (OPP) intensity histogram of representative siRNA-treated samples pulsed with OPP for 30 min and OPP-negative control (e). Quantification of mean fluorescent OPP intensity (f). OPP incorporation in siRNA-treated cortex cells in G₀G₁ (DNA content = 2c) and S/G₂/M (DNA content > 2c) phases of the cell cycle (g, h). DNA content was determined using propidium iodide. i–l Reduced protein translation rate in Yy1cKO cells at E12.5. OP-puro (OPP) intensity histogram of representative E12.5 control and Yy1cKO cells pulsed with OPP for 30 min and OPP-negative control (i). Quantification of mean fluorescent OPP intensity (j). OPP incorporation in cortical cells in G₀G₁ (DNA content = 2c) and S/G₂/M (DNA content > 2c) phases of the cell cycle (k, l). DNA content was determined using propidium iodide. Comparisons were performed using the two-tailed unpaired Student’s t test. Data are the mean ± standard deviation (c, f, h, j, l) and ± standard error of the mean (a, b). *p < 0.05. ns = not significant