Fig. 5: Perturbation of the cell cycle induced by glyoxal.

a HUVEC were treated with GO as indicated and subjected to cell cycle analysis applying a triple staining method (5-ethynyl-2′-deoxyuridine (EdU), p-H3 (S10), 4′,6-diamidino-2-phenylindole (DAPI)). Ratio of cells in the respective cell cycle phases (left) and dot plots representative for the 24 h-values (right) are shown. n = 4. b HUVEC were stained with carboxyfluorescein succinimidyl ester (CFSE) and treated with GO. CFSE staining was quantified by flow cytometry. n = 4. c Cell cycle proteins were chosen according to Reactome annotation divided by their specific role in each phase. Only proteins with significantly altered expression (absolute fold change > 0.58 and q < 0.05) or modified by CML are shown. Each dot represents the protein fold change and the black edge signifies proven CML modification. n = 4. d GO-treated HUVEC were lysed and subjected to immunoblot analysis. n = 5 (4 h), n = 6 (24 h). e Following GO treatment of HUVEC (8 h), immunofluorescence staining of γH2AX (S139) and DAPI was performed. Hydroxyurea (HU, 2 mM, 1 h) served as positive control. Representative pictures and quantification of γH2AX (S139)-positive nuclei are shown. Ctrl: control. Scale bar = 50 µm. n = 4. f HUVEC were treated with GO, lysed and subjected to immunoblot analysis. Densitometric quantification is shown in Supplementary Fig. 6c. n = 6. g HUVEC were treated with GO and intracellular (dichlorodihydrofluorescein diacetate (H2DCFDA)) and mitochondrial ROS (MitoSOX) were measured by flow cytometry. n = 4. a, b, d, e, g. Bar graphs and the curve diagram show mean + SEM. Statistical significance was analyzed using one-way or two-way repeated measurement ANOVA corrected via Holm–Šidák method. For panel e the positive control (HU) was not included into ANOVA. * p < 0.05 vs. respective control. Source data are provided as a Source Data file. Specific p values are listed in Supplementary Data 6. Related to Supplementary Fig. 6 and Supplementary Data 5.