Figure 6
SOCS6-mediated mitochondrial fragmentation is dependent on DRP1 fission activity. (a–d) AD293 cells were transfected with vectors encoding MFN1-myc, MFN2-myc, or DRP1-K38A alone or with pcDNA3-SOCS6. Immunofluorescent staining was performed after labeling cells with MitoTracker. Representative confocal images of HA-SOCS6 (blue, anti-SOCS6), MitoTracker, and MFN1-myc (green, anti-myc) (a) or DRP1-K38A (green, anti-DRP1) (c), are displayed at the bottom, whereas the top displays the merged, enlarged view. Scale bar: 10 μm. (b and d) Transfected cells were scored according to their mitochondrial morphology as tubular networks, hyperfused, or fragmented patterns. Percentages of cells with designated mitochondrial patterns are indicated in the bar graph. A total of 300 cells were counted for each category and data were derived from three independent experiments. (e–g) HeLa cells stably harboring shRNAs targeting luciferase (control-KD) and DRP1 (DRP1-KD) are shown. (e) DRP1 expression was determined by western blot analysis. (f) Confocal images of MitoTracker and DRP1 (green, anti-DRP1) or SOCS6 (green, anti-SOCS6) are shown in upper or bottom panel. Scale bar: 10 μm. (g) Transfected cells were scored according to their mitochondrial morphology as tubular networks, hyperfused, or fragmented patterns, and the percentages of distribution are shown in the bar graph. A total of 300 cells were counted for each category and data were derived from three independent experiments
