Journal: iScience

Article Title: A novel mitochondrial microprotein reprograms cellular bioenergetics and protects against hepatic steatosis

doi: 10.1016/j.isci.2026.117457

Figure Lengend Snippet: MNP33 is a novel mitochondrial inner-membrane protein (A) Workflow for identifying noncoding RNA-encoded proteins on LDs. The flowchart is adopted and modified from the figure in our previous work. , (B and C) Localization of transiently expressed MNP33. GFP control or MNP33-GFP was transiently transfected into HeLa cells. Then cells were viewed using a confocal microscope after being stained with MitoTracker Red for mitochondria or LipidTOX Red for LDs. Scale bars, 5 μm. (D) Diagram illustrates the locus and translation of MNP33. The nucleotide and amino acid sequences of MNP33 are shown (using BioEdit) (upper image). In mice, MNP33 (red) is located in exon 10 of the Cdhr4 gene (black) on chromosome 9 in Mouse GRCm39. MNP33 is translated in the +1 frame (lower image). (E) Localization of stably expressed MNP33. HeLa cells stably expressing GFP or MNP33-GFP were viewed using a confocal microscope after being stained with MitoTracker Red for mitochondria. Scale bars, 5 μm. (F) Distribution of MNP33 assessed by subcellular fractionation. HeLa cells stably expressing GFP or MNP33-GFP were treated with 100 μM OA for 12 h, and then the cells were fractionated. The proteins in different fractions were analyzed by western blotting, probing with the antibodies indicated. 33, MNP33-GFP; MT, mitochondria; TM, total membrane; PNS, post-nuclear supernatant. (G) Analysis of localization of MNP33 in mitochondrial membrane by proteinase K (PK) digestion assay. Mitochondria were isolated from HeLa cells stably expressing MNP33 and digested with PK at 37°C at the indicated concentrations. Western blotting analysis was performed to detect marker proteins of the mitochondrial outer membrane (CPT1A), inner membrane (Tim23), and MNP33. MNP33 was detected using antibodies against GFP or MNP33, respectively.

Article Snippet: Tim23 , ABclonal , Cat# A8688; RRID: AB_2772611.

Techniques: Membrane, Modification, Control, Transfection, Microscopy, Staining, Stable Transfection, Expressing, Fractionation, Western Blot, Isolation, Marker

Journal: iScience

Article Title: A novel mitochondrial microprotein reprograms cellular bioenergetics and protects against hepatic steatosis

doi: 10.1016/j.isci.2026.117457

Figure Lengend Snippet:

Article Snippet: Tim23 , ABclonal , Cat# A8688; RRID: AB_2772611.

Techniques: Virus, Expressing, Recombinant, Electron Microscopy, Protease Inhibitor, Bicinchoninic Acid Protein Assay, ATP Assay, TUNEL Assay, Plasmid Preparation, Software, Transfection