Experimental validation of computationally predicted gene enrichments.(A) Bar plot showing the log2 gene enrichments for the MB fraction-enriched MitoCarta 2.0 genes detected as highly and moderately enriched. (B) qRT-PCR validation of MB fraction-enriched MitoCarta 2.0 genes. (C) Log2 gene enrichments for the HS fraction-enriched MitoCarta 2.0 genes encoding mitochondrial ribosomal proteins, Mia40 substrates and TA proteins. (D) qRT-PCR validation of genes enriched in the HS fraction. Relative enrichment of genes in MB with respect to HS upon normalization with ERCC-00096 is shown. Data derived from three biological replicates. Error bars correspond to SEM; P < 0.05 (*), P < 0.01 (**), P < 0.001 (***), and P < 0.0001 (****) by unpaired t test. (E) Fluorescent in-situ hybridization of selected candidates by RNAscope on whole mount 5 days post fertilization zebrafish larvae. mRNAs are probed by specific RNAscope probes (red) and mitochondria are stained with anti-GFP antibody (green). Yellow in the merged image indicates co-localization of mRNAs on MLS-EGFP tagged mitochondria. Scale = 5 μm. (F) Analysis of co-localization of MB fraction-enriched MitoCarta 2.0 mRNAs with mitochondria. M1 indicates Mander’s co-localization co-efficient of mRNA fraction co-localized on mitochondria harboring MLS-EGFP. The difference between myod1 negative control and mRNA candidates were analyzed by unpaired t test. Error bars correspond to SEM and data derived from five region of interests originating from three experiments (n = 5); P < 0.0001 (****).
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