Supplementary material
for
Cytokinetic abscission is an acute G1 event
Ofir Gershony; Tal Pe’er; Meirav Noach; Natalie Elia; Amit Tzur*
Supplementary legends
Figure S1:
Live-cell imaging of HeLa cells stably expressing mKO2-Cdt1 undergoing a
complete cell cycle. Plot indicates mKO2-Cdt1 levels as a function of time. Cdt1
levels (fluorescence intensity) are provided as percentages of maximum level, i.e., the
plateau levels of mKO2-Cdt1. Measurements were taken every 15 min. Time 0 was
set at anaphase. Representative images from the movie sequence are shown below. n
= 10. Scale bar = 10 µm.
Figure S2:
BALB/c 3T3 cells (a) or HeLa cells (b) stably expressing mKO2-Cdt1 were
transfected with GFP-tubulin and imaged during late cytokinesis at 7-min intervals.
Time 0 was set at intercellular bridge-cutting. Graphs indicate the relative mKO2Cdt1 levels in the two newly formed daughter cells (red and orange lines), which did
not reach a 100% plateau in mKO2-Cdt1 levels within the time course of our
experiment. Percentages of mKO2-Cdt1 levels are relative to the highest intensity
value obtained throughout the time course of the experiment. Microtubule diameter
was measured based on the intensity profile of a line crossing the bridge connecting
the cells at the narrowest point along the bridge. Arrows indicate the beginning of
abscission, as characterized by an acute decrease in microtubule diameter at the first
1
abscission site. Notably, a considerable increase in mKO2-Cdt1 levels was observed
at abscission onset, as indicated by the acute reduction in microtubule diameter.
Movie S1:
Live-cell imaging of BALB/c 3T3 cells stably expressing mKO2-Cdt1 undergoing a
complete cell cycle imaged at 15-min intervals. Sum intensity projection overlaid on
DIC images is shown. Arrow indicates the dividing cell that was used for the mKO2Cdt1 measurements presented in Figure 1a.
Movie S2:
BALB/c 3T3 cells stably expressing mKO2-Cdt1 were transfected with GFP-tubulin
and imaged during late cytokinesis at 7-min intervals. Maximum intensity projections
are shown.
Movie S3:
BALB/c 3T3 cells stably expressing mKO2-Cdt1 were transfected with GFP-tubulin
and imaged during late cytokinesis at 7-min intervals. Maximum intensity projections
are shown.
Movie S4:
HeLa cells stably expressing mKO2-Cdt1 were transfected with GFP-tubulin and
imaged during late cytokinesis at 7-min intervals. Maximum intensity projections are
shown.
Movie S5:
2
HeLa cells stably expressing mKO2-Cdt1 were transfected with GFP-tubulin and
imaged during late cytokinesis at 7-min intervals. Maximum intensity projections are
shown.
3
Supplementary Figures
mKO2-Cdt1 (% of max)
Figure S1
Time (min)
Anaphase
0
2h
4h
6h
8h
10h
12h
14h
16h
18h
Figure S2
mKO2-Cdt1 (%)
Microtubule diameter (%)
BALB/c 3T3
100
80
60
40
20
0
-60 -40 -20
0
20 40
60 80
mKO2-Cdt1 (%)
Microtubule diameter (%)
HeLa
100
80
60
40
20
0
-60 -40 -20
0
20 40
4
60 80
20h
22h
24h
26h