Fig. 4: LSD1 inhibition suppresses HR and increases DNA DSBs. | Nature Communications

Fig. 4: LSD1 inhibition suppresses HR and increases DNA DSBs.

From: Repression of LSD1 potentiates homologous recombination-proficient ovarian cancer to PARP inhibitors through down-regulation of BRCA1/2 and RAD51

Fig. 4

a–d Representative images (a, c) and quantification (b, d) of neutral comet assays in A2780, SKOV3 and ES2 cells treated with indicated ZY0511 for 48 h or LSD1 knockdown (shLSD1) treatment after 5 Gy ionizing radiation (IR) treatment. Scale bar, 100 μm. Data represent mean ± SEM (unpaired two-tailed Student’s t test). The experiments were repeated three times. e, f Representative images (e) and quantification (f) of γH2AX-foci staining performed in A2780, SKOV3 and ES2 cells with or without 1 μM LSD1i (ZY0511 or SP2577) for 48 h or LSD1 knockdown (shLSD1) treatment. Green, γH2AX; blue, DAPI. Scale bar, 10 μm. Data represent mean ± SEM of three biologically independent experiments (unpaired two-tailed Student’s t test). g, h Representative images (g) and quantification (h) and of RAD51 nuclear foci in A2780, SKOV3 and ES2 cells with or without 1 μM LSD1i (ZY0511 or SP2577) for 48 h or LSD1 knockdown (shLSD1) treatment at 4 h after 2 Gy IR treatment. Green, RAD51; blue, DAPI. Scale bar, 10 μm. Data represent mean ± SEM of three biologically independent experiments (unpaired two-tailed Student’s t test). i–l Quantification of γH2AX foci and RAD51 foci per nucleus at the indicated time points after 2 Gy IR treatment in ES2 cells treated with or without 1 μM LSD1i (ZY0511 or SP2577) for 48 h (i, k), or shRNA suppression of LSD1 (j, l). Data represent mean ± SEM of three biologically independent experiments (two-way ANOVA). m Schematic illustration of the GFP-based HR reporter assay (DR-GFP) and NHEJ reporter assay (EJ5-GFP). iGFP, internal GFP repeat. n, o Quantification of HR and NHEJ using DR-GFP and EJ5-GFP reporter assay, respectively. Data represent mean ± SEM of three biologically independent experiments (unpaired two-tailed Student’s t test). ns, not significant, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001. Source data and exact p values are provided as the Source Data file.

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