KDM2A/B–RNF2 axis is necessary for H2AUb signaling and DNA damage response. (A) Quantification of total KDM2A, KDM2B, and RNF2 protein levels in control knockdown (siCTRL), in KDM2A and RNF2-depleted cells (siKDM2A + siRNF2) or in KDM2B and RNF2-depleted cells (siKDM2B + siRNF2) with or without etoposide treatment. Blot shows two biological replicates. Protein levels were normalized to loading control (GAPDH). (B) Representative immunofluorescence images of H2AUb signal (gray) within siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells either left untreated or treated briefly with 10 μM etoposide. Scale bar = 20 μm. Images are representative of three independent experiments conducted on separate days. N ≥ 100 nuclei imaged. (C) Quantification of nuclear H2AUb signal intensity in siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells with or without etoposide treatment. Dots represent the number of nuclei measured. Nuclei are indicated for representative experiment and N ≥ 100 nuclei per condition. (D) Representative immunofluorescence images of 53BP1 (green) within siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells either left untreated or treated briefly with 10 μM etoposide. Scale bar = 20 μm. Images are representative of two independent experiments conducted on separate days. N ≥ 100 nuclei imaged. (E) Quantification of the number of 53BP1 foci per nucleus in siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells with or without etoposide treatment. Nuclei are indicated for representative experiment and N ≥ 50 nuclei per condition. (A, C, and E) Data represent means ± SD. Statistical differences between groups were analyzed employing one-way ANOVA Tukey’s multiple comparison test between each group and a control. Source data are available for this figure: SourceData F8.
Figure 8.

KDM2A/B–RNF2 axis is necessary for H2AUb signaling and DNA damage response. (A) Quantification of total KDM2A, KDM2B, and RNF2 protein levels in control knockdown (siCTRL), in KDM2A and RNF2-depleted cells (siKDM2A + siRNF2) or in KDM2B and RNF2-depleted cells (siKDM2B + siRNF2) with or without etoposide treatment. Blot shows two biological replicates. Protein levels were normalized to loading control (GAPDH). (B) Representative immunofluorescence images of H2AUb signal (gray) within siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells either left untreated or treated briefly with 10 μM etoposide. Scale bar = 20 μm. Images are representative of three independent experiments conducted on separate days. N ≥ 100 nuclei imaged. (C) Quantification of nuclear H2AUb signal intensity in siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells with or without etoposide treatment. Dots represent the number of nuclei measured. Nuclei are indicated for representative experiment and N ≥ 100 nuclei per condition. (D) Representative immunofluorescence images of 53BP1 (green) within siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells either left untreated or treated briefly with 10 μM etoposide. Scale bar = 20 μm. Images are representative of two independent experiments conducted on separate days. N ≥ 100 nuclei imaged. (E) Quantification of the number of 53BP1 foci per nucleus in siCTRL cells, siKDM2A + siRNF2 cells, or siKDM2B + siRNF2 cells with or without etoposide treatment. Nuclei are indicated for representative experiment and N ≥ 50 nuclei per condition. (A, C, and E) Data represent means ± SD. Statistical differences between groups were analyzed employing one-way ANOVA Tukey’s multiple comparison test between each group and a control. Source data are available for this figure: SourceData F8.

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