Fig. 4: Mitochondria protect 1C metabolism-dependent serine catabolism upon cytosolic 1C pathway inhibition. | Nature Communications

Fig. 4: Mitochondria protect 1C metabolism-dependent serine catabolism upon cytosolic 1C pathway inhibition.

From: Mitochondria preserve an autarkic one-carbon cycle to confer growth-independent cancer cell migration and metastasis

Fig. 4

a Cytosolic and mitochondrial 1C metabolism and the pharmacologic intervention points of MTX. b Absolute consumption and release (CORE) rates of serine (Ser) and glycine (Gly) from MDA-MB-468 cells upon 24 h 50 nM MTX. Dots represent the mean of an individual experiment each measured in triplicate wells; mean ± SEM (n = 6). c Formate release rate upon [U-13C]serine in MDA-MB-468 cells after 24 h 50 nM MTX; mean ± SEM of three independent experiments each measured in triplicate wells; unpaired t-test with Welch’s correction for M + 1 isotopologue (n = 3). d MID of intracellular serine upon [U-13C]serine in MDA-MB-468 cells upon 24 h 50 nM MTX; mean ± SEM of four independent experiments each measured in triplicate wells. e Ratio of absolute intracellular serine and glycine after 24 h 50 nM MTX in MDA-MB-468 cells. Dots represent the mean of an independent experiment in triplicate wells; mean ± SEM (n = 5). f MID of intracellular serine upon [U-13C]serine in SHMT1, MFT, SHMT2, and FPGS-depleted HAP1 cells; mean ± SEM of independent experiments each measured in triplicate wells (n = 8 for CTRL, SHMT1ko, and SHMT2ko; n = 5 for MTX; n = 2 for FPGSko, MFTko and MFT/SHMT1ko). g Ratio of absolute intracellular serine and glycine levels in SHMT1, MFT, SHMT2, and FPGS depleted HAP1 cells. Dots represent the mean of an independent experiment in triplicate wells; mean ± SEM (n = 8 for CTRL and SHMT2ko, n = 7 for SHMT1ko, n = 6 for MTX, n = 2 for FPGSko, MFTko and MFT/SHMT1ko). h Absolute CORE rates of formate from WT and SHMT1- or SHMT2-depleted HAP1 cells upon 24 h 50 nM MTX. Each dot indicates the mean of an individual experiment measured in triplicate wells; Brown-Forsythe and Welch ANOVA test with Dunnetts multiple comparisons test. Mean ± SEM (n = 4 for CTRL, MTX, SHMT2ko, SHMT2ko MTX; n = 3 for SHMT1ko and SHMT1ko MTX) i Absolute CORE rates of formate from WT and MTHFD1L-depleted MDA-MB-468 cells upon 24 h 50 nM MTX. Mean ± SEM (n = 6 for CTRL and MTX; n = 3 for MTHFD1Lko + /- MTX) of an individual experiment measured in triplicate wells; Brown-Forsythe and Welch ANOVA test with Dunnetts multiple comparisons test. j Absolute CORE rates of formate from SCR and MTHFD2 KD MDA-MB-468 cells upon 24 h 50 nM MTX. Dots represent mean of an individual experiment measured in triplicate wells; mean ± SEM; (n = 3) Brown-Forsythe and Welch ANOVA test with Dunnetts multiple comparisons test. k Absolute CORE rates of formate from MDA-MB-468 cells upon 24 h treatment with 2.5 mM Metformin and 50 nM MTX. Dots represent mean of an individual experiment measured in triplicate wells; mean ± SEM (n = 2). Source data are provided as a Source Data file.

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