Figure 6.
Lattice light sheet imaging of Spindly dynamics in live cells. (a) Video stills of siControl-treated HeLa cells expressing doxycycline-inducible LAP-Spindly after nocodazole washout. Cells were arrested in 3.3 µM nocodazole for 4 h and washed once, and a full cell volume was acquired using a lattice light sheet microscope every 1 min for 1 h. Insets show zooms of single kinetochores from the detection panel. Scale bar, 5 µm. (b) Video stills of siCENP-F–treated HeLa cells expressing doxycycline-inducible LAP-Spindly after nocodazole washout. Cells were arrested in 3.3 µM nocodazole for 4 h and washed once, and a full cell volume was acquired using a lattice light sheet microscope every 1 min for 1 h. Insets show zooms of single kinetochores from the detection panel. Scale bar, 5 µm. (c) Quantification of the cumulative LAP-Spindly intensity for all kinetochores at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Error bars are ±SD. (d) Quantification of the spot number at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Errors bars are ±SD. (e) Quantification of the median spot intensity at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Errors bars are ±SD.

Lattice light sheet imaging of Spindly dynamics in live cells. (a) Video stills of siControl-treated HeLa cells expressing doxycycline-inducible LAP-Spindly after nocodazole washout. Cells were arrested in 3.3 µM nocodazole for 4 h and washed once, and a full cell volume was acquired using a lattice light sheet microscope every 1 min for 1 h. Insets show zooms of single kinetochores from the detection panel. Scale bar, 5 µm. (b) Video stills of siCENP-F–treated HeLa cells expressing doxycycline-inducible LAP-Spindly after nocodazole washout. Cells were arrested in 3.3 µM nocodazole for 4 h and washed once, and a full cell volume was acquired using a lattice light sheet microscope every 1 min for 1 h. Insets show zooms of single kinetochores from the detection panel. Scale bar, 5 µm. (c) Quantification of the cumulative LAP-Spindly intensity for all kinetochores at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Error bars are ±SD. (d) Quantification of the spot number at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Errors bars are ±SD. (e) Quantification of the median spot intensity at each time point over the 60-min video for siControl- and siCENP-F–treated cells. Measurements were made using TrackMate in Fiji. Errors bars are ±SD.

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