Figure S3.
Depletion of Top2α does not impair actin-patch formation in cytokinesis with chromatin bridges. (A and B) Actin patches (empty arrowheads) in cytokinesis with chromatin bridges in BE cells, in the absence (control) or presence of Top2α siRNA. The bases of the canals are shown at higher magnification. Broken chromatin bridges are indicated by dotted arrows. (C) Relative actin-patch intensity values. (D) Mean intensity of INCENP at the midbody ring in cells with chromatin bridges. Values in control were set to 1. Mean ± SD from n cells. (E–I) Western blot analysis of total Rad50, ATM, Nbs1, Chk2, INCENP, Aurora B, actin, and tubulin. (J) Experimental procedure for Top2-inhibition by ICRF-193 or etoposide in cells enriched in cytokinesis after release from a nocodazole block. (K and L) Mean intensity of Mre11 or ATM at the bridge DNA next to the midbody in cells with chromatin bridges after nocodazole release, in the absence (control) or presence of 10 μM ICRF-193 or 10 μM etoposide. (M) Mean intensity of INCENP to the midbody ring in cells with chromatin bridges after nocodazole release. Mean ± SD from n cells. Values in control were set to 1. (N) Frequency of broken chromatin bridges in cytokinesis after nocodazole release. Mean ± SD from three independent experiments (n > 120). (O–Q) Mean intensity of Mre11 or ATM at the bridge DNA next to the midbody and of Aurora B at the midbody ring in HU-induced BE cells with chromatin bridges. Mean ± SD from n cells. Values in control were set to 1. ***, P < 0.001 (ANOVA and Student’s t test). Numbers below/next to each bar indicate n. Values in control were set to 1. (R–T) Localization of Top2α and Rad17 at the midbody in normally segregating cells. Tubulin values indicate midbody thickness. Midbodies (boxed areas) were reexported and are shown at higher magnification. Scale bars, 5 μm. Source data are available for this figure: SourceData FS3.

Depletion of Top2α does not impair actin-patch formation in cytokinesis with chromatin bridges. (A and B) Actin patches (empty arrowheads) in cytokinesis with chromatin bridges in BE cells, in the absence (control) or presence of Top2α siRNA. The bases of the canals are shown at higher magnification. Broken chromatin bridges are indicated by dotted arrows. (C) Relative actin-patch intensity values. (D) Mean intensity of INCENP at the midbody ring in cells with chromatin bridges. Values in control were set to 1. Mean ± SD from n cells. (E–I) Western blot analysis of total Rad50, ATM, Nbs1, Chk2, INCENP, Aurora B, actin, and tubulin. (J) Experimental procedure for Top2-inhibition by ICRF-193 or etoposide in cells enriched in cytokinesis after release from a nocodazole block. (K and L) Mean intensity of Mre11 or ATM at the bridge DNA next to the midbody in cells with chromatin bridges after nocodazole release, in the absence (control) or presence of 10 μM ICRF-193 or 10 μM etoposide. (M) Mean intensity of INCENP to the midbody ring in cells with chromatin bridges after nocodazole release. Mean ± SD from n cells. Values in control were set to 1. (N) Frequency of broken chromatin bridges in cytokinesis after nocodazole release. Mean ± SD from three independent experiments (n > 120). (O–Q) Mean intensity of Mre11 or ATM at the bridge DNA next to the midbody and of Aurora B at the midbody ring in HU-induced BE cells with chromatin bridges. Mean ± SD from n cells. Values in control were set to 1. ***, P < 0.001 (ANOVA and Student’s t test). Numbers below/next to each bar indicate n. Values in control were set to 1. (R–T) Localization of Top2α and Rad17 at the midbody in normally segregating cells. Tubulin values indicate midbody thickness. Midbodies (boxed areas) were reexported and are shown at higher magnification. Scale bars, 5 μm. Source data are available for this figure: SourceData FS3.

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