Figure 3.

Detached Cells Have Altered Mitochondrial Morphology and Function

(A) Representative transmission electron microscopy images of mitochondria of attached and detached cells. Scale bars represent 1 μm. Graphs present quantified cristae length, cristae/outer membrane ratio and number of cristae per mitochondria (Att n = 58 mitochondria and Det n = 283 mitochondria). Data are presented as mean ± SD. ^∗∗∗∗p ≤ 0.0001, unpaired t test.

(B) Intracellular levels of unlabeled and M + 4-labeled aspartate from ¹³C₅-labeled glutamine in attached and detached HeLa and 293T cells. Peak area levels are normalized to cell number.

(C) Oxygen consumption rate profiles of 293T cells cultured in attached or detached conditions. Dotted lines show incubation of cells in the presence of indicated mitochondrial inhibitors. Data are normalized to protein content.

(D) Mitochondrial ROS production in attached and detached 293T cells as measured by MitoSOX Red staining and analysis by flow cytometry.

(B–D) Data are presented as mean ± SD of triplicate cultures of representative experiments.