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. Author manuscript; available in PMC: 2014 Oct 14.
Published in final edited form as: Cancer Cell. 2013 Oct 3;24(4):450–465. doi: 10.1016/j.ccr.2013.08.020

Figure 3. Glutamine Restriction Induces S-phase Stalling in a Subset of Basal TNBC.

Figure 3

Icon codes in figure keys. Icons represent mean values +/−SD. (A) Growth curves of “Glutamine Sensitive” carcinomas (underlined in Figure 2A) in glutamine-free media. (B) Percent increases in Annexin V reactivity of cells in glutamine-free media at day 3. Group averages; A, 0.8%,+/−1.4; B 1.2% +/−1.4; C, 5.8% +/−3.2; t-test A vs C p= 0.005. (C) Paired bars representing the change in percent of cells in G2/M (Δ% G2/M) with nocodazole treatment at day 5 in control (gray) versus glutamine-free (yellow) media, using cell cycle curve fitting software (FLOJO, Treestar). (D) Paired bars representing the percent S-phase fraction with nocodazole treatment of day 5 cultures in control (gray) vs. glutamine-free (yellow) media. (E) S phase stalling is accompanied by a decrease in total and Serine 780 phosphorylated retinoblastoma protein. (F) Culture confluence reduces S-phase fractions and glutamine sensitivity; S-phase decrease (Δ%), is the decrease in % S-phase with high density culture (gray icons) versus low density culture (colored icons); paired icons connected by dashed lines represent a single cell line. (G) Doubling time (from Figure 1C) vs. glutamine-free culture sizes (from Figure 2H). See also Figure S3.