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Figure 4.
Figure 4. Cdc42 Activation Is Not Required for TJ
Assembly(A) MDCK cells expressing myc-Cdc42 (T17N) under control
of doxycycline (DC) were grown ± 20 ng/ml DC till
confluence and then subjected to calcium switch. Cells were
fixed 6 hr later and stained for myc-epitope and ZO-1.(B)
Transient expression of the CRIB domain of γPAK has no effect
on TJs. Cells were fixed and stained for the myc epitope and
ZO-1 24 hr posttransfection.(C) Par6ΔPro[136] does not
coimmunoprecipitate Cdc42. COS cells were transfected with
plasmids encoding HA[3]-Cdc42 (Q61L) and/or myc-Par6B or
myc-Par6BΔPro[136] as indicated. Par6B was immunoprecipitated,
and bound Cdc42 was detected by immunoblot. CL = cell lysate
(1/10 of total).(D) Effect of myc-Par6BΔPro[136] on TJ
formation.(E) Percentage of cells lacking complete TJs between
cells that express myc-Par6B or myc-Par6BΔPro[136] at 6 hr
postcalcium addition.
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