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Tao Lin and Jasneet Kaur
Lab Advisor: Dr. Edward Bonder
Teaching Assistant: Susan Seipel
Is there a correlation
between spindle orientation in
mitotic cells and the contracted
cell shape caused by the
GTPase protein, RhoA?
RhoA
mDiaRho kinase
Myosin phosphatase
LimK
Cofilin
Actin-myosin filament stabilization
MLC-p MLC
Actin polymerization
Unbranched filaments
Stress fibers
Actin-myosin contractility
(cell rounding)
EB1/APC
Microtubule growth and stabilization
Spread, control
cell
Contracted cell with
constitutively active RhoA
Note: These images have been generated through a 90 degree vertical projection of z-series images
Y27632 treatment
• Y27632 inhibits Rho kinase which causes actin-myosin contractility leading to rounded cell shape.
• Different concentrations of Y27632 used 0μM, 1.0 μM, 1.5 μM, 2.0 μM 2.5 μM, 5.0 μM, 10.0 μM
• High concentrations of Y27632 allow the cell shape to return to normal
• Goal: What is the point where cell shape remains contracted but the spindle orientation returns to normal? – Will show that the spindle misorientation is not a
direct effect of contracted shape caused by RhoA, and thus, is caused by a different aspect of the RhoA signaling pathway.