Fig. 3: Hypoxia improves the anti-proliferative capacity of C2C12 myotubes against EO771 cells. | Oncogenesis

Fig. 3: Hypoxia improves the anti-proliferative capacity of C2C12 myotubes against EO771 cells.

From: Transcriptomic profiling of co-cultured cancer-host cells identifies hypoxia as a driver of the skeletal muscle cell’s anti-proliferative effect on cancer cells

Fig. 3

A HIF-1α protein level in normoxic and hypoxic (6 h at 3% O2) conditions in monocultures and co-cultures. The full Western blot membrane images are shown in Fig. S7. B Schematic representation of monocultures. C Extracellular lactate concentration (n = 7), D Extracellular glucose concentration (n = 3) and E Medium pH measurements performed at room temperature (n = 2, each replicate represents pooled supernatants from 3 to 4 wells of a 12-well plate) in supernatants from monocultures of EO771 (40,000 cells per well seeded in 12-well plate at D0), MLg and C2C12 (80,000 cells per well seeded in 12-well plate) after 2 days of incubation under normoxic or hypoxic conditions. F Schematic representation of co-cultures. G Outgrowth of EO771 alone and EO771 in co-cultures with either C2C12 myotubes (C2C12 + EO771) or MLg cells (MLg + EO771) under normoxic or hypoxic conditions over 2 days. Experiments were performed in 96-well plates. EO771 area represents the percentage of fluorescent cells normalized to day 0 (D0) (n = 34–40). Representative fluorescence microscopy images of EO771 (4000 cells per well in 96-well plate) cell outgrowth on C2C12 or MLg at day 2 (D2). Scale bar = 200 µm. H Outgrowth of EO771 (4000 cells per well in 96-well plate) in co-cultures with C2C12 myotubes under normoxic or hypoxic conditions over 2 days. EO771 area represents the percentage of fluorescent cells normalized to D0 (n = 30–36). I Extracellular lactate concentration (n = 8), J Extracellular glucose concentration (n = 3–4), and K Medium pH measurements performed at room temperature (n = 2, each replicate represents pooled supernatants from 20 wells of a 96-well plate) in supernatants from co-cultures under normoxic or hypoxic conditions after 2 days. Data are mean ± SEM. In each figure, samples were collected from two independent experiments (except glucose). Comparison between groups was done using two-way ANOVA with Šidák (only two groups) or Tukey’s (more than two groups) comparisons. **P < 0.01; ****P < 0.0001.

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