LBL1 induced DSBs in cancer cells. (A) LBL1 arrested the cells at G2/M phase. The cell cycle distribution
of
MDA-MB-231 cells treated with different concentrations of LBL1 for 48 h. Then the cell cycle profile was analyzed by flow cytometry
after the cells were fixed and stained with PI. (B) LBL1 induced G2/M arrest and G1 peak broadening in MDA-MB-231 cells.
The cells were treated with LBL1 for 48 h. The different
cell cycle stages (G1, S, G2/M) are indicated. (C) LBL1 did not induce phosphorylation of H2AX in normal primary HFF. The
cells were treated with indicated drugs for 24 h. Then the cells were
collected, and the lysates were prepared for Western blot analysis
with indicated antibodies. CPT was used as a positive control. (D,
E) Quantification of γ-H2AX-foci-positive MDA-MB-231 cells.
Data are presented as mean ± SD (n = 3, ∼100
cells were analyzed for each experimental condition). Representative
fluorescence micrographs are shown in part D, and quantification is
shown in part E. (F) Quantification of tail DNA from the neutral COMET
assay in MDA-MB-231 cells treated with LBL1. Around 200
cells were analyzed for each condition (n = 193 for
DMSO and n = 203 for LBL1-treated cells).
The representative micrographs are shown in Figure S3B.