Figure 10. CD2AP, JACOP, and CapZ are functional components of the SH3BP1 complex. (A) siRNA-transfected A431 cells were stimulated with EGF and then stained with fluorescent phalloidin. Shown are epifluorescence images of the apical (0 and 5 min) and of the basal (30 min) side of the cells. Note that CD2AP- and JACOP-depleted cells form neither dorsal ruffles nor a cortical actin belt. (B) siRNA-transfected A431 cells were serum starved and then stimulated with EGF for different periods of time. The cells were then fixed and stained for ZO-1. Shown are epifluorescence images taken from the apical part of the cells. Note that depletion of SH3PB1 and its associated proteins prevents the formation of ZO-1–positive ruffles and junctional recruitment. (C) Caco-2 cells transfected with siRNAs targeting SH3BP1, CD2AP, and JACOP were processed for immunofluorescence with antibodies against the three proteins. Shown are epifluorescence images of the junctional region. (D) Levels of active Cdc42 were measured after serum starvation and stimulation with EGF in extracts from cells transfected with either control, CD2AP-, or JACOP-targeting siRNAs. Shown are means ± 1 SD (n = 4). (E) A431 cells transfected with control and SH3BP1-targeting siRNAs and, after serum starvation, treated for 5 min with EGF. After fixation, the cells were stained as indicated. Shown are epifluorescence images representing dorsal focal planes. (F) A431 cells transfected with siRNAs targeting CapZα1 were then analyzed by immunoblotting as indicated. (G) A431 cells were siRNA transfected, serum starved, and then stimulated with EGF. The cells were then fixed and stained with fluorescent phalloidin. Shown are epifluorescence images. Note that depletion of CapZα1 leads to induction of filopodia and loss of the junctional actin belt. Bars, 10 µm.
Figure 10.

CD2AP, JACOP, and CapZ are functional components of the SH3BP1 complex. (A) siRNA-transfected A431 cells were stimulated with EGF and then stained with fluorescent phalloidin. Shown are epifluorescence images of the apical (0 and 5 min) and of the basal (30 min) side of the cells. Note that CD2AP- and JACOP-depleted cells form neither dorsal ruffles nor a cortical actin belt. (B) siRNA-transfected A431 cells were serum starved and then stimulated with EGF for different periods of time. The cells were then fixed and stained for ZO-1. Shown are epifluorescence images taken from the apical part of the cells. Note that depletion of SH3PB1 and its associated proteins prevents the formation of ZO-1–positive ruffles and junctional recruitment. (C) Caco-2 cells transfected with siRNAs targeting SH3BP1, CD2AP, and JACOP were processed for immunofluorescence with antibodies against the three proteins. Shown are epifluorescence images of the junctional region. (D) Levels of active Cdc42 were measured after serum starvation and stimulation with EGF in extracts from cells transfected with either control, CD2AP-, or JACOP-targeting siRNAs. Shown are means ± 1 SD (n = 4). (E) A431 cells transfected with control and SH3BP1-targeting siRNAs and, after serum starvation, treated for 5 min with EGF. After fixation, the cells were stained as indicated. Shown are epifluorescence images representing dorsal focal planes. (F) A431 cells transfected with siRNAs targeting CapZα1 were then analyzed by immunoblotting as indicated. (G) A431 cells were siRNA transfected, serum starved, and then stimulated with EGF. The cells were then fixed and stained with fluorescent phalloidin. Shown are epifluorescence images. Note that depletion of CapZα1 leads to induction of filopodia and loss of the junctional actin belt. Bars, 10 µm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal