Fig. 1
From: APC/CCDH1 synchronizes ribose-5-phosphate levels and DNA synthesis to cell cycle progression
Cell cycle-coupled TKTL1 expression regulates R5P levels. a Protein levels of TKTL1, TKT, and TKTL2 were determined in double thymidine-synchronized G1/S and RO3306-synchronized G2/M HeLa cells. Synchronizing effects were detected by flow cytometry (see Supplementary Fig. 1). b, c Protein levels of TKTL1, TKT, TKTL2, RPIA, CDC20, and CDH1 were determined at different time points after HeLa cells were released from (b) double thymidine and (c) RO3306 synchronization. Quantitative results of TKTL1 are shown below. Values are means ± SEM of three independent experiments. The cell cycle phases of indicated time points were confirmed by flow cytometry (see Supplementary Fig. 1). d R5P levels in HeLa cells at different cell cycle phases were measured after cells were released from double thymidine (upper panel) and RO3306 (lower panel) synchronization. Cell phases were confirmed by flow cytometry sorting. Data are presented by means ± SEM of five independent experiments. e R5P levels in both HEK293T and HeLa cells as well as TKT or TKTL1-overexpressing HEK293T and HeLa cells were determined. Data are shown as average of five independent experiments, presented as means ± SEM, two-tailed Students’ t test, **p < 0.01, ns not significant. f R5P levels in both HEK293T and HeLa cells as well as TKTL1-knockdown HEK293T and HeLa cells were determined. Values are shown as average of five independent experiments, presented as means ± SEM, two-tailed Students’ t test, *p < 0.05, ns not significant vs. control group. g Overexpression of TKTL1 in ccRCC. Representative immunohistochemical staining (IHC) and quantitative results of 12 samples are shown. T, tumor tissue; N, adjacent non-cancer tissue. Scale bars: 200 μm. Data are presented as means ± SEM and two-tailed Students’ t test was used, ***p < 0.001. h Expression levels of TKTL1, TKT, RPIA, and CDH1 in ccRCC. Protein levels of ccRCC tumors and matched adjacent non-cancer tissues were analyzed by western blotting (left). Quantitative results (n = 24 pairs of ccRCC tumors tissues and adjacent tissues; right) are shown as means ± SEM, Student’s t test, ***p < 0.001, **p < 0.01, *p < 0.05, ns not significant. i Average R5P concentrations were determined for 24 paired ccRCC tumors and their matched non-cancer tissues. Data are shown as means ± SEM, Student’s t test, ***p < 0.001. Full-length blots are presented in Supplementary Fig. 10
