Extended Data Fig. 5: Impact of NADK2 and mtFAS on mitochondrial translation.
(a) Coomassie staining of purified ACP-FLAG from WT and ΔMECR HeLa cells, which was then subjected to Asp-N peptidase treatment for assessment of acyl-ACP species via LC/MS. (b) Representative immunofluorescence images for ACP-FLAG (green) detected with anti-FLAG antibody MitoTracker Red (red), and nuclei (blue) stained with Hoechst for WT and ΔNADK2 HeLa cells cultured in the presence of 0.2 mM proline. Scale bars, 10 µm. (c) The NADK2 coessentiality network showing the top 6 enrichment gene clusters. (d) The MECR coessentiality network showing the top 6 enrichment gene clusters. (e) Volcano plot illustrating the log2 fold change of mitoribosome-related proteins identified in ACP immunoprecipitates from wild-type (WT), ΔNADK2, and ΔMECR HeLa cells. Proteins belonging to the mitochondrial ribosomal protein large subunit (MRPL) or small subunit (MRPS), as well as other interactors, are indicated and color-coded. (f) Schematic illustrating the workflow of click chemistry-based mitochondrial translation assay.
