![Figure 4. ALIX and TSG101 cooperatively control abscission timing. (A) Live imaging of HeLa cells stably expressing Histone2B-mCherry and eGFP–α-tubulin upon siRNAs treatment as indicated. Cumulative frequency plot showing the time interval between furrow ingression and abscission (n > 100 cells from three independent experiments; control, 74 ± 17.3 min; ALIX siRNA, 177 ± 78.7 min; TSG101 siRNA, 98 ± 28.4 min; ALIX + TSG101 siRNA, 338.2 ± 228.9 min [± SD]; ALIX relative to control, P < 0.001; TSG101 relative to control, P = 0.04; ALIX+TSG101 relative to ALIX, P = 0.002; ALIX+TSG101 relative to all others, P < 0.001; P-values obtained using mixed factor model). (B) Knockdown efficiency of ALIX and TSG101. (C) Live imaging of HeLa cells stably expressing Histone2B-mCherry, eGFP–α-tubulin, and CHMP4B-V5 or CHMP4BΔALIX-V5 upon siRNA treatment as indicated. Cumulative frequency plot showing the time interval between furrow ingression and abscission (n > 100 cells from three independent experiments; CHMP4B: control, 82 ± 19.9 min; CHMP4B siRNA, 74 ± 14.5 min; CHMP4B+TSG101 siRNA, 110 ± 25.6 min; CHMP4BΔALIX control, 80 ± 24.0 min; CHMP4B siRNA, 84 ± 24.7 min; CHMP4B+TSG101 siRNA, 153 ± 50.4 min [± SD]; CHMP4B with control siRNA or CHMP4B siRNA relative to CHMP4B with CHMP4B+TSG101 siRNA, P < 0.05; CHMP4BΔALIX with CHMP4B+TSG101 siRNA relative to all others, P < 0.001; P-values obtained using mixed factor model). (D) Knockdown efficiency of CHMP4B and TSG101 as well as expression of CHMP4B-V5 or CHMP4BΔALIX-V5 (*, nonspecific immunoreactivity).](https://cdn.rupress.org/rup/content_public/journal/jcb/212/5/10.1083_jcb.201507009/6/jcb_201507009_fig4.jpeg?Expires=1771639983&Signature=WSZ4usXyled1HFSD5OJIhPNrFDf5YuGmJpA33nNkzAbiA1o8hkXg5SPDN77g6-W0U3NM7T7cRwCC7vx1Aoa0u7jUIw~BrZX6f~qIJDCeCgow-g~VGYIFYNlCcCKDmf4EFtrUl3YlQ1~ySyMoW8gawjuygB8MBYaFCmgfvsZFgdc5gZEKONbWyz1hr22Ulq5NHGbfsu7K9z7Li3PmpIENUxVemybHoG5DxNnU4~Wk3qf31OA-onIUgkBMnECbLb77d-ruo997DZQAdzYDpoPeBX1DMNwbWkHAh~~24KBzoFRTkXZNQtjLpzQWobFT0tFpkFbRxsZHEqJ6EqYEfdh2pg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
ALIX and TSG101 cooperatively control abscission timing. (A) Live imaging of HeLa cells stably expressing Histone2B-mCherry and eGFP–α-tubulin upon siRNAs treatment as indicated. Cumulative frequency plot showing the time interval between furrow ingression and abscission (n > 100 cells from three independent experiments; control, 74 ± 17.3 min; ALIX siRNA, 177 ± 78.7 min; TSG101 siRNA, 98 ± 28.4 min; ALIX + TSG101 siRNA, 338.2 ± 228.9 min [± SD]; ALIX relative to control, P < 0.001; TSG101 relative to control, P = 0.04; ALIX+TSG101 relative to ALIX, P = 0.002; ALIX+TSG101 relative to all others, P < 0.001; P-values obtained using mixed factor model). (B) Knockdown efficiency of ALIX and TSG101. (C) Live imaging of HeLa cells stably expressing Histone2B-mCherry, eGFP–α-tubulin, and CHMP4B-V5 or CHMP4BΔALIX-V5 upon siRNA treatment as indicated. Cumulative frequency plot showing the time interval between furrow ingression and abscission (n > 100 cells from three independent experiments; CHMP4B: control, 82 ± 19.9 min; CHMP4B siRNA, 74 ± 14.5 min; CHMP4B+TSG101 siRNA, 110 ± 25.6 min; CHMP4BΔALIX control, 80 ± 24.0 min; CHMP4B siRNA, 84 ± 24.7 min; CHMP4B+TSG101 siRNA, 153 ± 50.4 min [± SD]; CHMP4B with control siRNA or CHMP4B siRNA relative to CHMP4B with CHMP4B+TSG101 siRNA, P < 0.05; CHMP4BΔALIX with CHMP4B+TSG101 siRNA relative to all others, P < 0.001; P-values obtained using mixed factor model). (D) Knockdown efficiency of CHMP4B and TSG101 as well as expression of CHMP4B-V5 or CHMP4BΔALIX-V5 (*, nonspecific immunoreactivity).
