Figure 5. CB002 and its analogs perturbed an S-phase rather than a G2-phase checkpoint like other known xanthines cell cycle effects in SW480 cells.

Western blot analysis of synchronized SW480 treated cells as indicated and harvested at 24 hr (A, B, C). Synchronized SW480 cells were treated as indicated and analyzed by Western blot (D), PI staining (E) or PI/BrdU analysis (F). CB002 (C), Caffeine (CF). Experiments from figures (A)–(D) were performed at least twice and a representation of one is shown.

Figure 5—figure supplement 1. Flow cytometry PI/BrdU-CB002-analog #4 perturbs the S-phase rather than the G2 checkpoint, unlike other xanthines.

Synchronized SW480 cells were treated as indicated, chased with BrdU for 30 min, and harvested over a time course of 0–48 hr. Cells were double-stained for Propidium Iodide and BrdU (A, B). Haploid cell gatings indicate the haploid BrdU-positive cells.