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. 2022 Jan 29;298(3):101649. doi: 10.1016/j.jbc.2022.101649

Figure 9.

Figure 9

Functional evaluation of IMP-1-bound mRNAs.A–C, mitochondrial function is impaired in IMP-1-knockdown cells. Mitochondrial activity was assayed in either mouse EP cells (A) or human breast cancer cells (MCF-7 cells; B) using Seahorse assay. About 16,500 EP and 12,500 MCF-7 cells were used for each assay shown (representative of three assays). Assays were run either with complete media (with 25 mM glucose and 4 mM glutamine) or with media with only one of these carbon sources, as indicated. ∗∗p < 0.05; ∗∗∗p < 0.001. C, MitoTracker staining of EP cells confirmed a reduced mitochondrial membrane potential (MitoΨ) in IMP1-knockdown cells (shown for two shRNAs). D and E, glutathione peroxidase (GPX) mRNAs were reduced in IMP1-knockdown cells. Relative (mRNA) was assayed for IMP1-binding mRNA species GPX-1 and GPX-2 in EP cells after (partial) knockdown of IMP-1, alongside the mitochondrial ribosomal protein Mrpl14 mRNA, also pulled through in association with IMP-1 (D). Western blotting confirmed that GPX-2 protein was also depleted (E). ECAR, extracellular acidification rate; EP, epithelial; IMP-1, insulin-like growth factor 2 mRNA-binding protein-1; OCR, oxygen consumption rate.