Figure 1. Chemical Screen Identifies Glutamine Metabolism as a Vulnerability in SIRT3 KO Cells.

(A) Layout of experimental design for the small-molecule screen.

(B) Primary screen data highlighting the top five compounds inhibiting SIRT3 KO cell growth (see also Figures S1A–S1D and Table S1).

(C) Schematic of the structures of glutamine and its analogs, azaserine and 6-diazo-5-oxo-L-norleucine (DON).

(D and E) Dose-response curves of WT and SIRT3 KO MEF growth after treatment with azaserine (D) or DON (E) for 72 hr.

(F) Cell viability measured by propidium iodide incorporation in WT and SIRT3 KO MEFs after 72 hr of glutamine deprivation.

(G) Relative growth of WT and SIRT3 KO MEFs treated with 30 μM azaserine compared to control for 4 days.

(H) Clonogenic growth assay of Kras (G12V)-transformed WT and SIRT3 KO MEFs treated with DMSO (as a control) or 30 mM azaserine for 8 days.

(I) Quantification of colony formation assay using ImageJ (NIH) to calculate the area of each well covered by WT or SIRT3 KO MEF colonies (n = 3).

(J and K) Glutamine uptake (J) and ammonia production (K) in WT or SIRT3 KO MEFs after 24 hr (see also Figures S1E–S1K).

Data in this figure are represented as mean ± SEM. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.