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. 2009 Jul 6;58(10):2303–2315. doi: 10.2337/db07-1781

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© 2009 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 6. — Fis1 RNAi restores mitochondrial morphology and dynamics under HFG in INS1 cells. A: Mitochondrial morphometry. Mitochondria were classified according to AR into short (AR <2), intermediate, (2<AR<4), and long (AR >4) length (n = 8 cells per group). B: Whole-cell mitochondrial fusion assays (PAGFPmt dilution) in Fis1 RNAi INS1 cells exposed to HFG for 24 h (▲). A hyperbolic fitting yielded T₅₀ = 7.6 min. Fis1 RNAi cells not exposed to HFG are also plotted (♦). Values of control RNAi cells with and without HFG treatment were imported from Fig. 3C (■ and ○, respectively). C: Mitochondrial movement was calculated by measuring the velocity of single mitochondria over time. Velocities are not significantly different between groups (P >0.05). Control RNAi with and without HFG treatment are represented by ■ and □, respectively. Fis1 RNAi cells with and without HFG treatment are represented by and ▤, respectively. D: Confocal images demonstrating mitochondrial movement analysis. Images at time 0 and 10 min are portrayed. The last image zooms to one mitochondrion and the movement over 10 min is depicted, in this case 4 μm over 10 min. The velocity over time is plotted and indicates a peak around 250 s. When the distance from the origin is plotted over time, it is evident that mitochondria do not move at a constant speed. (A high-quality digital representation of this figure is available in the online issue.)

Inline graphic — Fis1 RNAi restores mitochondrial morphology and dynamics under HFG in INS1 cells. A: Mitochondrial morphometry. Mitochondria were classified according to AR into short (AR <2), intermediate, (2<AR<4), and long (AR >4) length (n = 8 cells per group). B: Whole-cell mitochondrial fusion assays (PAGFPmt dilution) in Fis1 RNAi INS1 cells exposed to HFG for 24 h (▲). A hyperbolic fitting yielded T₅₀ = 7.6 min. Fis1 RNAi cells not exposed to HFG are also plotted (♦). Values of control RNAi cells with and without HFG treatment were imported from Fig. 3C (■ and ○, respectively). C: Mitochondrial movement was calculated by measuring the velocity of single mitochondria over time. Velocities are not significantly different between groups (P >0.05). Control RNAi with and without HFG treatment are represented by ■ and □, respectively. Fis1 RNAi cells with and without HFG treatment are represented by and ▤, respectively. D: Confocal images demonstrating mitochondrial movement analysis. Images at time 0 and 10 min are portrayed. The last image zooms to one mitochondrion and the movement over 10 min is depicted, in this case 4 μm over 10 min. The velocity over time is plotted and indicates a peak around 250 s. When the distance from the origin is plotted over time, it is evident that mitochondria do not move at a constant speed. (A high-quality digital representation of this figure is available in the online issue.)