Figure 5.

DPC Removal during S-Phase of the Cell Cycle

(A) Schematic of the experimental approach used to synchronize T24 cell in G0 and monitor DPC levels during S-phase entry.

(B) Total DPC levels in G0-, late G1-, and S-phase before and after SPRTN depletion in HeLa cells visualized by silver staining. The WB of cyclin A and cell-cycle profiles were used as a control of S-phase entry.

(C) WB showing SPRTN expression or depletion in non-replicative and replicative T24 cells.

(D) Schematic of cell viability measurement in replicative and non-replicative cells.

(E) Cell viability of replicative and non-replicative T24 cells after SPRTN depletion and FA treatment.

(F) Total DPC levels after G1/S release in HeLa WT and Δ-SPRTN cells visualized by silver staining. The WB of cyclins and cell-cycle profiles (lower images) were used to control S-phase progression.

(G) WB showing presence of DNA-PK and Lamin B1 in DPCs in Δ-SPRTN HeLa cells after G1/S release. Whole cell extract: WCE. Mean ± SEM, n = 3.