Fig. 4: Glyoxal impairs the proliferation of HUVEC. | Nature Communications

Fig. 4: Glyoxal impairs the proliferation of HUVEC.

From: Mapping protein carboxymethylation sites provides insights into their role in proteostasis and cell proliferation

Fig. 4

a Left, CM-modified proteins annotated to different cellular compartments according to Gene Ontology annotation after GO treatment (1 mM, 48 h). ECM: extracellular matrix; ER: endoplasmic reticulum. Right, Gene Ontology cellular component terms enriched in CM-modified proteins. False Discovery Rate (FDR) < 0.05. b Left, Volcano plot depicting proteins that significantly increase (red) or decrease (blue) abundance upon GO treatment (1 mM, 48 h) or remain unchanged (gray). Horizontal dashed line indicates a significance cut-off of q < 0.05 and vertical dashed lines an absolute fold change (log2) > 0.58. n = 4. Right, Gene set enrichment analysis for Gene Ontology biological process terms based on protein fold changes. Terms enriched among increased (red) or decreased (blue) proteins are shown. FDR < 0.05; NES: normalized enrichment score. c-i. HUVEC were treated with GO for 48 h. c Cell numbers were determined. n = 6. d Cells were stimulated with basic fibroblast growth factor (bFGF) (50 ng/ml, 24 h) and BrdU incorporation was measured. n = 5. e Spheroids were generated, embedded, and stimulated with vascular endothelial growth factor (VEGF) (50 ng/ml, 24 h). Representative pictures and sprout numbers per spheroid are shown. Scale bar=100 µm. n = 4. f, g Oxygen consumption rate (OCR) (f) and extracellular acidification rates (ECAR) (g) were measured via Seahorse technology and mitochondrial and glycolytic parameters calculated. n = 6. h Cells were stained with MitoTracker and analyzed by flow cytometry. n = 5. i Intracellular ATP levels were measured in cell extracts. n = 3. c–i Data are represented as mean + SEM. Statistical significance was analyzed using one-way or two-way repeated measurement ANOVA corrected via Holm–Šidák method. * p < 0.05 vs. control, # p < 0.05 vs. respective non-bFGF (d) or non-VEGF-treated sample (e). Source data are provided as a Source Data file. Specific p values are listed in Supplementary Data 6. Related to Supplementary Fig. 5 and Supplementary Data 4.

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