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. Author manuscript; available in PMC: 2019 Aug 27.
Published in final edited form as: Nature. 2019 Feb 27;568(7750):117–121. doi: 10.1038/s41586-019-0977-x

Figure 2. Pyruvate drives availability of α-ketoglutarate which metabolically regulates P4HA activity.

Figure 2

(a) Hydroxylated collagen based on hydroxyproline (OH-proline) in MCF10A H-RASV12 spheroids in the presence or absence of pyruvate in combination with cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM), citrate (5 mM), or malate (5 mM).

(b) Relative change in hydroxylated collagen based on hydroxyproline (OH-proline) in human (MCF10A H-RASV12, MCF7) and mouse (4T1, EMT6.5) breast cancer spheroids transduced with lentiviral CRISPR with or without guide for MCT2 upon addition of cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM) in the presence of pyruvate. Data are normalized to the corresponding conditions without genetic MCT2 inhibition. Dashed line indicates the level of hydroxylated collagen in the corresponding conditions without genetic MCT2 inhibition.

(c) Hydroxylated collagen based on hydroxyproline (OH-proline) in human (MCF10A H-RasV12, MCF7, HCC70) and mouse (4T1) breast cancer spheroids in the presence of pyruvate upon addition of 1.5 mM (MCF10A H-RasV12, 4T1) or 2 mM (MCF7, HCC70) cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG) and/or 1.5 mM cell permeable succinate (dimethyl succinate).

Error bars represent SD of mean from biological independent samples (n=3). Two-tailed unpaired student’s T-test.