Figure 4.

Detached Cells Are Dependent on Glycolysis for ATP Production and Survival

(A) Cell viability of 293T, HeLa, and A549 cells cultured in attached and detached conditions in the presence or absence of glucose. Cells were cultured in attached or detached conditions for 7 days in the presence of glucose before glucose starvation for 24 h followed by staining with eFlour 780 viability dye and analysis by flow cytometry.

(B) Intracellular ATP and AMP levels of 293T attached and detached cells in the presence or absence of glucose. Cells were grown in attached and detached conditions like in (A) before glucose was removed for 4 h followed by LC-MS analysis.

(C) Cell viability of 293T cells as measured in (A) in cells cultured with glucose or without glucose plus indicated ATP concentrations.

(D and E) Levels of M + 2 citrate (D) and other TCA cycle intermediates (E) originating from ¹³C₁₆-palmitate in 293T attached and detached cells cultured with or without glucose. Cells were cultured as in (A) before addition of labeled palmitate and removal of glucose for 4 h.

(F) Confocal microscopy images of attached and detached HeLa cells expressing Mito YFP and stained with Bodipy C11.

(A, B, D, and E) Data are represented as mean ± SD of triplicate cultures of representative experiments.