Fig. 2
From: Mitochondria facilitate neuronal differentiation by metabolising nuclear-encoded RNA
Acquisition of neRNAs by mitochondria facilitates emergence of neuronal transcriptome during differentiation. (A) Quantification of mRNAs of interest in mitochondrial fractions isolated from differentiating NPCs at different time points (as per the X axis). Values are normalised to levels of the corresponding transcripts in control NPCs at baseline (T0). * two-tailed p-value < 0.0001, error bars: ±1SD, n = 3 biological replicates. (B) Quantification of mRNAs of interest in mitochondrial fractions isolated from differentiating PNPaseRNAi NPCs at different time points (as per the X axis). Values are normalised to levels of the corresponding transcripts in PNPaseRNAi NPCs at baseline (T0). * two-tailed p-value < 0.0001, error bars: ±1SD, n = 3 biological replicates. (C) MA plot shows Log2 profile of neRNAs acquired by mitochondria isolated from the differentiating cells at t = 1 h relative to mitochondrial neRNAs isolated from the control NPCs (T0). (D) Line graphs show the intensity of SYBR Green I dye (y-axis) used in gel electrophoresis of RNAs isolated from mitochondria and nuclei of differentiating NPCs at different time points, plotted against molecular weight of RNAs (x-axis). (E) Left photomicrograph shows polyadenylated neRNAs identified using a FISH probe in control (T0) and differentiating (1 h: T1) NPCs in vitro. Right micrograph shows polyadenylated neRNAs identified using a FISH probe in ventricular and subventricular zone of a rostral region of E13 mouse neural tube. Note a mosaic pattern of expression characterised by a diminished level of polyA+ neRNAs in the subventricular zone cells. Left scale bars top: 8 μm, bottom: 2 μm. Right scale bar: 30 μm. (F) MA plot shows Log2 profile of neRNAs isolated from the nuclei of differentiating NPCs at t = 1 h and 2 h relative to nuclear neRNAs of the control NPCs (T0). (G) Top bar plot shows expression level of Ngn1 in PNPaseRNAi differentiating NPCs normalised to control differentiating NPCs. Bottom bar plots demonstrate expression level of PNPase in RNAiPnpt1-treated versus control NPCs at baseline and after induction of differentiation. * two-tailed p-value < 0.0001, n = 3 biological replicates