Journal: Scientific Reports
Article Title: A cleavage product of Polycystin-1 is a mitochondrial matrix protein that affects mitochondria morphology and function when heterologously expressed
Figure Lengend Snippet: Recombinant PC1 localizes to mitochondria in a subset of cells. ( a . The CTF can be further processed to release a cytoplasmic tail (CTT). Constructs used in the study include full-length human PC1 (hFL) with N-terminal (eGFP or mCherry) and C-terminal (HA or eGFP) tags. ( b ) Lysates from MDCK cells with either stable, inducible expression of vector control (pcDNA5) or eGFP-PKD1-HA constructs probed with GFP or HA antibodies show that the majority of cellular PC1 is free NTF, with only a small amount of uncleaved FL and CTF. ( c – f ) NIH3T3 cells transfected with mCherry-PKD1-eGFP show three main patterns: (1) NTF in the ER with undetectable CTF ( c ); (2) FL and/or NTF/CTF either mostly co-localized in the ER ( d ) or with partial co-localization in the ER and with CTF/CTT in distinct subcellular regions ( e , panel insert); (3) NTF localized to the ER and CTF or CTT in mitochondria ( f ). The endoplasmic reticulum (ER) is identified by transient expression of pEF.myc.ER-E2-Crimson or stained with ER-Tracker™ Blue-White DPX ( c , d ) and mitochondria (Mito) are identified by transient expression of mito-BFP or stained with MitoTracker Deep Red.
Article Snippet: Pkd1 inactivation was confirmed using genomic PCR and/or reverse-transcriptase PCR (TaqMan gene expression assay, Applied Biosystems, cat. no. 4351372, Mm00465436_g1).
Techniques: Recombinant, Construct, Expressing, Plasmid Preparation, Transfection, Staining