![Figure 8. Loss of AFG3L2 induces aberrant accumulation of MT-ATP6. (A) Immunoblot of HEK293 whole cell lysates after AFG3L2 knockdown with two independent siRNAs compared with scrambled control (sc). The asterisk indicates unspecific signal detected with the AFG3L2 antibody. (B) Immunoblotting of sucrose density gradient fractions of AFG3L2 siRNA. The asterisks indicate unspecific signal in fractions 1 and 2 detected only with MRPS18b antibody with TCA precipitated samples. (C, left) A 30-min 35S metabolic labeling of cells in A. The arrow indicates MT-ATP6. (right) quantification of the signal intensity of MT-ATP6 relative to MT-CO1 in the same sample for two independent biological experiments of #2 AFG3L2 siRNA and scrambled control (sc). (D) HEK293 cells treated with the indicated siRNA metabolically labeled for 45 min with [35S] methionine/cysteine in the presence of anisomysin and then precipitated with CTAB. Each sample was split equally in two with one half treated with RNase A before addition of CTAB. A representative image of independent biological experiments is shown. a.u., arbitrary units.](https://cdn.rupress.org/rup/content_public/journal/jcb/211/2/10.1083_jcb.201504062/6/jcb_201504062_fig8.jpeg?Expires=1773575024&Signature=GKbdrPw~4NwLfKmElluIX6o5aWzZ21Pz~aNUOL6Lcu2CmWPllhNKOUssVm-LoLMxyOU6Y3bnO3~EDd5xH1Hm5HBdDKQepKS6w~P7XdaX~U~CZaxKLyYFKtGew6GxIXxdoeE1zpWxiq3BJXgLNHC-5krk8DnQufR9UC0ejMraKVCu9-K~OqHNOMm348kXS2QO8CekIUYqpgpUYIJ4PHom2AUrqch5UZOWOhV7vwi2afWLqyeUJpNlYenKCnpyYYN-08aVb~AcFKxweId2zqL5wxxLx81IffAlPDAO9jKlERyA47ZUeCkMhAI~SNV3dS68UTm3AAb9NS3TSAl1z~S5wA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Loss of AFG3L2 induces aberrant accumulation of MT-ATP6. (A) Immunoblot of HEK293 whole cell lysates after AFG3L2 knockdown with two independent siRNAs compared with scrambled control (sc). The asterisk indicates unspecific signal detected with the AFG3L2 antibody. (B) Immunoblotting of sucrose density gradient fractions of AFG3L2 siRNA. The asterisks indicate unspecific signal in fractions 1 and 2 detected only with MRPS18b antibody with TCA precipitated samples. (C, left) A 30-min 35S metabolic labeling of cells in A. The arrow indicates MT-ATP6. (right) quantification of the signal intensity of MT-ATP6 relative to MT-CO1 in the same sample for two independent biological experiments of #2 AFG3L2 siRNA and scrambled control (sc). (D) HEK293 cells treated with the indicated siRNA metabolically labeled for 45 min with [35S] methionine/cysteine in the presence of anisomysin and then precipitated with CTAB. Each sample was split equally in two with one half treated with RNase A before addition of CTAB. A representative image of independent biological experiments is shown. a.u., arbitrary units.
