Skip to main content
. Author manuscript; available in PMC: 2026 Jan 31.
Published in final edited form as: ACS Chem Biol. 2025 Dec 19;21(1):26–36. doi: 10.1021/acschembio.5c00639

Figure 4. Analysis of FANCD2 and γH2AX foci formation in HeLa cells treated with colibactin 686 (9).

Figure 4.

γH2AX (gray) and FANCD2 (blue) foci formation quantified by percent of cells with ≥10 γH2AX foci or ≥5 FANCD2 foci in HeLa cells treated with various doses of colibactin 686 (9) is shown. Conditions: HeLa cells were incubated with colibactin 686 (9, 0.08–15000 nM) for 4 h, washed, cultured an additional 20 h, fixed with paraformaldehyde and immunostained using FANCD2 and γH2AX primary antibodies and fluorescently labeled secondary antibodies. Foci were visualized using ImageXpress Micro 4 high content imager (20× magnification, 9 fields of view acquired), and mean number of foci per nucleus as well as % positive cells were quantified using Metaxpress image analysis software. DNA damage foci in cells treated with DMSO vehicle control or olaparib and MMC positive controls were quantified using the same conditions. Columns indicate the mean; n ≥ 6 technical replicates (individual values are shown).