Figure S4.
Comt depletion results in acentric failure to enter nuclei and nuclear envelope reassembly defects.(A and B) Stills from time-lapse videos of mitotic neuroblasts expressing H2Av-RFP (magenta), GFP-NLS (green), I-CreI with wild-type levels of Comt (A), or with RNAi-depleted levels of Comt (B). In control neuroblasts (A), acentrics (arrows) segregate poleward and enter daughter nuclei. Nuclei recruit GFP-NLS (arrowheads) while acentrics are still separate from nuclei. In Comt-depleted neuroblasts (B), acentrics segregate but fail to enter daughter nuclei, forming micronuclei (arrowheads). Nuclei recruit GFP-NLS while acentrics are still separate from nuclei, although GFP-NLS signal is not as strong or evenly distributed as it is in nuclei from control neuroblasts. Time is written in seconds after initial acentric poleward movement. Scale bars are 2 µm. (C) GFP-NLS intensity on daughter nuclei from control neuroblasts (black line; n = 22) and neuroblasts expressing RNAi against Comt (purple line; n = 30). Lines represent averages. Dark-shaded regions represent ± the standard error. Light-shaded regions represent ± twice the standard error. Dashed lines indicate time of nuclear envelope reassembly initiation. On average, nuclear envelope reassembly initiated at a comparable rate in control neuroblasts and neuroblasts with Comt RNAi. However, GFP-NLS intensity plateaued on nuclei from Comt-depleted divisions compared with control divisions, resulting in overall less signal intensity. Dashed green line represents average time that acentrics began to enter channels (155 s). (C′) GFP-NLS intensity on neuroblasts expressing RNAi against Comt where acentrics reentered daughter nuclei (blue line; n = 15) or where acentrics formed micronuclei (pink line; n = 15). GFP-NLS signal intensity was lower on daughter nuclei from Comt-depleted neuroblasts when acentrics formed micronuclei than from Comt-depleted neuroblasts when acentrics reentered nuclei. Dashed green line represents average time that acentrics began to enter channels (155 s). Difference was determined to not be statistically significant (P = 0.17) by a Scheirer–Ray–Hare test. See also Fig. 5.

Comt depletion results in acentric failure to enter nuclei and nuclear envelope reassembly defects. (A and B) Stills from time-lapse videos of mitotic neuroblasts expressing H2Av-RFP (magenta), GFP-NLS (green), I-CreI with wild-type levels of Comt (A), or with RNAi-depleted levels of Comt (B). In control neuroblasts (A), acentrics (arrows) segregate poleward and enter daughter nuclei. Nuclei recruit GFP-NLS (arrowheads) while acentrics are still separate from nuclei. In Comt-depleted neuroblasts (B), acentrics segregate but fail to enter daughter nuclei, forming micronuclei (arrowheads). Nuclei recruit GFP-NLS while acentrics are still separate from nuclei, although GFP-NLS signal is not as strong or evenly distributed as it is in nuclei from control neuroblasts. Time is written in seconds after initial acentric poleward movement. Scale bars are 2 µm. (C) GFP-NLS intensity on daughter nuclei from control neuroblasts (black line; n = 22) and neuroblasts expressing RNAi against Comt (purple line; n = 30). Lines represent averages. Dark-shaded regions represent ± the standard error. Light-shaded regions represent ± twice the standard error. Dashed lines indicate time of nuclear envelope reassembly initiation. On average, nuclear envelope reassembly initiated at a comparable rate in control neuroblasts and neuroblasts with Comt RNAi. However, GFP-NLS intensity plateaued on nuclei from Comt-depleted divisions compared with control divisions, resulting in overall less signal intensity. Dashed green line represents average time that acentrics began to enter channels (155 s). (C′) GFP-NLS intensity on neuroblasts expressing RNAi against Comt where acentrics reentered daughter nuclei (blue line; n = 15) or where acentrics formed micronuclei (pink line; n = 15). GFP-NLS signal intensity was lower on daughter nuclei from Comt-depleted neuroblasts when acentrics formed micronuclei than from Comt-depleted neuroblasts when acentrics reentered nuclei. Dashed green line represents average time that acentrics began to enter channels (155 s). Difference was determined to not be statistically significant (P = 0.17) by a Scheirer–Ray–Hare test. See also Fig. 5.

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