Figure 3
From: Human Mitochondrial DNA-Protein Complexes Attach to a Cholesterol-Rich Membrane Structure
Knock-down of ATAD3 disrupts the cholesterol containing fraction and with it the membrane associated nucleoid.
Pellet preparations from digitonin treated purified mitochondria derived from ATAD3 siRNA treated HEK293 cells were separated on a bottom-up flotation iodixanol gradient. The cholesterol and corresponding nucleoid components no longer migrate tightly as one fraction, but are spread evenly over two fractions. mtDNA, mitochondrial DNA; polG, polymerase Gamma; SSB, single-stranded binding protein; TFAM, mitochondrial transcription factor A; PHB, prohibitin, MRPS, mitochondrial ribosomal protein small subunit; MRPL, mitochondrial ribosomal protein large subunit; Clnx, calnexin; coxII, cytochrome oxidase subunit II; TOM20, translocase of outer membrane 20 kDa subunit; VDAC1, voltage-dependent anion channel 1. Protein, DNA and lipid profiles of replicate gradients are presented in SFig. 2.
