Fig. 3: MTHFD2 is required for increased cellular glycine levels downstream of TGF-β. | Nature Communications

Fig. 3: MTHFD2 is required for increased cellular glycine levels downstream of TGF-β.

From: Mitochondrial one-carbon metabolism is required for TGF-β-induced glycine synthesis and fibrotic responses

Fig. 3

a Schematic representation of metabolite labeling downstream of 2,3,3-D3-Serine. Human lung fibroblasts (HLFs) were transfected with siRNA targeting MTHFD2 or nontargeting siRNA. Cells were labeled with 2,3,3-D3-Serine and treated with TGF-β for 48 hours or left untreated. b–g Gas chromatography/mass spectrometry analysis of cellular metabolites. b Analysis of cellular glycine labeling from 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. c Relative levels of M + 1 glycine from (b). d Analysis of cellular serine after labeling with 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. e Relative levels of M + 1 and M + 2 serine from (d). f Analysis of cellular proline labeling from 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. g Relative levels of M + 1 and proline from (f). h Relative levels of media formate content in NHLFs treated with TGF-β or left untreated. i–k Liquid chromatography/mass spectrometry analysis of cellular metabolites. i Analysis of cellular GTP labeling from 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. j Analysis of cellular ATP labeling from 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. k Analysis of cellular SAM labeling from 2,3,3-D3-Serine in control and MTHFD2 knockdown HLFs. Bar graphs represent mean ± SD, n = 3 biological replicates (b–g, i–k), n = 4 biological replicates (h). Significance was calculated by 2-way ANOVA with Tukey’s post-test. Source data are provided as a Source Data file.

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