Figure 3.

AHNAK regulates 53BP1-mediated G1-S transition upon DSB induction in transformed and non-transformed cells

(A) Schematic representation of the G1 checkpoint assay. Briefly, 24 h post-transfection with the indicated siRNA, MCF7 cells were serum arrested in G₀ for 24 h and subsequently released in serum-containing medium containing nocodazole. At 4 h, cells were treated with IR (4 Gy) and collected at 0, 4, 16, and 22 h time points before examined by flow cytometry. Solid green bars denote experimental time points.

(B) Cell-cycle distributions were analyzed using flow cytometry in WT, 53BP1 ^−/−, and two independent AHNAK^−/− MCF7 cell lines, transfected with control or 53BP1 siRNA.

(C) Quantification of the relative distribution of cells in S-G2 phase at the experimental time points, relative to 0 h.

(D) Schematic representation of the G1 checkpoint assay for BJ fibroblast.

(E) Cell-cycle distributions were analyzed using flow cytometry in BJ cell lines, transfected with control or indicated siRNA.

(F) Quantification of the relative distribution of cells in S-G2 phase at the experimental time points, relative to 0 h.