Fig. 4

Acquisition of neRNAs reprograms mitochondrial metabolism. (A) Representative electron microscopy photomicrographs of NPCs at t = 0.5 h and 1 h after induction of differentiation. Note the presence of early (arrows) and late (arrowheads) autophagosomes (pseudo-coloured turquoise) in differentiating NPCs. Scale bars left: 2 μm, middle: 1 μm, right: 0.4 μm. (B) Graph shows the ATP content of NPCs subsequent to induction of neuronal differentiation. * two-tailed p-value < 0.0001, n = 3 biological replicates. (C) Graph shows profile of cellular metabolites of central carbon metabolome subsequent to induction of neural differentiation (n = 1 sample/time point). Y-axis values represent Log₂ (Area under the curve) for the studied metabolites at different time points during differentiation normalised to Log₂ (Area under the curve) for the metabolites at baseline. (D) Western blots show the impact of differentiation on downstream mediators of AMPK and the pro-neural transcription factor Neurogenin-2. (E) Bar plot shows expression level of Ngn1 in inhibitor-treated differentiating NPCs normalised to control differentiating NPCs. * two-tailed p-value < 0.0001. (F) Schematic demonstration of timeframe of events triggered by inter-organellar communication during differentiation of NPCs. (G) Bar plot shows the level of ATP in antimycin-A-treated NPCs receiving isolated and reprogrammed donor mitochondria normalised to the ATP level in antimycin-A-treated recipient NPCs at baseline. * two-tailed p-value < 0.001, n = 3 biological replicates