DSBs induce accumulation of PCM proteins at the centrosome. (A) Left: NEDD1, PCNT, and γ-tubulin were recruited to the centrosomes in G1 cells 4 h after IR. Right: Semiquantitative analysis of the relative immunostaining signal intensity of PCM proteins at the centrosomes in control or IR-treated U2OS cells was shown in box plot. Scale bar, 20 µm. (B) DMSR was determined in G1 cells treated with scrambled shRNA, shNEDD1, or shPCNT. Quantitation of microtubule length is shown (n > 50). (C) Semiquantitative analysis of relative PCNT signal intensity at the centrosomes in control or PLK1 inhibitor–treated G1 cells is shown in box plot (n > 50). (D) G1 cells were pretreated with PLK1 or P38 inhibitor for 30 min and exposed to 2 Gy IR. DMSR was determined 4 h after IR. Quantitation of microtubule length in control, PLK1 inhibitor, or p38 inhibitor treated cells is shown (n > 50). (E) Semiquantitative analysis of relative PCNT or NEDD1 signal intensity at the centrosomes from control or DNA-PK inhibitor–treated G1 cells is shown in box plot (n > 50). (F) Semiquantitative analysis of relative PCNT or NEDD1 signal intensity at the centrosomes in G1 cells treated with scrambled (NC) siRNA or si53BP1 is shown in box plot (n > 50). For box plot, center line, median; box limits, 25th and 75th percentile; whiskers, minimum and maximum (see also Materials and methods). ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant. A.U., arbitrary units; MT, microtubule; UT, untreated.
Figure 5.

DSBs induce accumulation of PCM proteins at the centrosome. (A) Left: NEDD1, PCNT, and γ-tubulin were recruited to the centrosomes in G1 cells 4 h after IR. Right: Semiquantitative analysis of the relative immunostaining signal intensity of PCM proteins at the centrosomes in control or IR-treated U2OS cells was shown in box plot. Scale bar, 20 µm. (B) DMSR was determined in G1 cells treated with scrambled shRNA, shNEDD1, or shPCNT. Quantitation of microtubule length is shown (n > 50). (C) Semiquantitative analysis of relative PCNT signal intensity at the centrosomes in control or PLK1 inhibitor–treated G1 cells is shown in box plot (n > 50). (D) G1 cells were pretreated with PLK1 or P38 inhibitor for 30 min and exposed to 2 Gy IR. DMSR was determined 4 h after IR. Quantitation of microtubule length in control, PLK1 inhibitor, or p38 inhibitor treated cells is shown (n > 50). (E) Semiquantitative analysis of relative PCNT or NEDD1 signal intensity at the centrosomes from control or DNA-PK inhibitor–treated G1 cells is shown in box plot (n > 50). (F) Semiquantitative analysis of relative PCNT or NEDD1 signal intensity at the centrosomes in G1 cells treated with scrambled (NC) siRNA or si53BP1 is shown in box plot (n > 50). For box plot, center line, median; box limits, 25th and 75th percentile; whiskers, minimum and maximum (see also Materials and methods). ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant. A.U., arbitrary units; MT, microtubule; UT, untreated.

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