![Figure 1. FAM92A1 is a ubiquitously expressed mitochondrial protein. (A) Western blot analysis of FAM92A1 expression in different cell lines. (B) Immunofluorescence staining of U2OS cells for endogenous FAM92A1. Bars: 10 µm (main images); 5 µm (insets). (C) Western blot analysis to examine the subcellular localization of FAM92A1. M, mitochondrial fraction (pellet [P]); R, rest fraction except mitochondria (the supernatant [S]) after final centrifugation at 7,000 g. (D) Protease digestion of isolated mitochondria with proteinase K. TOM20, cytochrome c, COXIV, and Hsp60 were used as mitochondrial protein controls in different submitochondrial compartments. (E) ImmunoEM of endogenous FAM92A1. Bars: 200 nm (main images); 50 nm (inset). The red arrows indicate FAM92A1. (F) Quantification of immunogold particle distribution in the MOM/IBM and CM. ***, P ≤ 0.001, Student's t test. (G) Mitochondrial protein extraction by sodium carbonate. T, total mitochondria. (H) Schematic diagram of FAM92A1 N-terminal peptides with different tags. (I) Immunofluorescence images of cells transfected with FAM92A11–47-GFP and FAM92A1Δ1–47-GFP for 24 h. Bars, 10 µm. (J) Transmission EM analysis of mitochondrial import of FAM92A11–47-Apex2. Bars, 200 nm.](https://cdn.rupress.org/rup/content_public/journal/jcb/218/1/10.1083_jcb.201806191/26/jcb_201806191_fig1.jpeg?Expires=1771608243&Signature=ssFtl-pOOWIkjPKecOmYo-4z1W1pBaTLIhMKpvxuNcyd45rEAEKqqZH5rz99gV~RiBwpQ5KFEeMCxNg1nQUNyFZR7bj1XVUNAHq~BoE7LcxZzX6sCbHeK6at9uNGPNwgAEGSpkCrNiv3NtSGjGvuH9a-eKR0Qld02Uhlvk6agZUxpPjsPPtgZx5WvtkXlW8V96Glzz~GBdOp~AVc6T2AIvaiYySQIJ9huDwaaWkMUu~45nP-Ab~G4Dqoo~3IntT6LohssmHJMFBIq8rEy8IYHoTzZEhum0DhJiTSB6XhVocCkdq1FDBeKNI59ejgM1lrXlBsr892k1k3ApusQRVX3g__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
FAM92A1 is a ubiquitously expressed mitochondrial protein. (A) Western blot analysis of FAM92A1 expression in different cell lines. (B) Immunofluorescence staining of U2OS cells for endogenous FAM92A1. Bars: 10 µm (main images); 5 µm (insets). (C) Western blot analysis to examine the subcellular localization of FAM92A1. M, mitochondrial fraction (pellet [P]); R, rest fraction except mitochondria (the supernatant [S]) after final centrifugation at 7,000 g. (D) Protease digestion of isolated mitochondria with proteinase K. TOM20, cytochrome c, COXIV, and Hsp60 were used as mitochondrial protein controls in different submitochondrial compartments. (E) ImmunoEM of endogenous FAM92A1. Bars: 200 nm (main images); 50 nm (inset). The red arrows indicate FAM92A1. (F) Quantification of immunogold particle distribution in the MOM/IBM and CM. ***, P ≤ 0.001, Student's t test. (G) Mitochondrial protein extraction by sodium carbonate. T, total mitochondria. (H) Schematic diagram of FAM92A1 N-terminal peptides with different tags. (I) Immunofluorescence images of cells transfected with FAM92A11–47-GFP and FAM92A1Δ1–47-GFP for 24 h. Bars, 10 µm. (J) Transmission EM analysis of mitochondrial import of FAM92A11–47-Apex2. Bars, 200 nm.
