Figure 2. THP-1 and TIME cells stably expressing wild-type K5 have an altered metabolism.

(A) Medium from equal numbers of the indicated cells were examined at 3 days post-seeding both visually (left panel) and for absorbance at 570 nm (O.D.₅₇₀) (right panel) (B) Equal numbers of each THP-1 cell line were starved for 3 hours and incubated in glucose free DMEM medium with a fluorescent glucose analog (2-NBDG) (0.1 mM) for a period of 30 minutes. Live cells were gated and the fluorescence intensity assessed by flow cytometry at the indicated time points. A representative experiment of analog uptake (left panel) and the quantification of fold change after 30 min. with standard deviations (right panel) are shown. (C) Cells were seeded in triplicate at 0.1 x 10⁶ cells/ml and supernatant was taken at the indicated time points and subjected to colorimetric measurement of lactate concentration. (D) Cells were seeded at 0.1 x 10⁶ cells/ml in glucose-free DMEM media supplemented with different concentrations of glucose (12 mM, 6 mM, 2 mM or 0 mM). Viable cell numbers were determined daily. Data are representative of three experiments. (E) Normalized whole cell lysate of each of the indicated THP-1 and TIME cell lines were subject to western blot (WB) using an anti-phospho-PKM2 (Tyr 105) antibody and re-probed with anti-PME-1 antibodies.