Figure 7. Endogenous RYK and MIB1 are required for Wnt/CTNNB1 signaling at the level of receptor activation. (A) RYK and MIB1 siRNA transfection inhibits cDNA overexpression-mediated activation of BAR by Wnt-3A and Disheveled (DVL3) but not CTNNB1 or nondegradable CTNNB1* (S33A, S37A, T41A, and S45A). Data are mean + SD (error bars). (B) Western blot of cell lysates after a time course of treatment with Wnt-3A–conditioned media in cells transfected with control or MIB1 siRNAs. (C) Western blot after 1 h of treatment with Wnt-3A–conditioned media in HEK293T cells transfected with the indicated siRNAs. In rows labeled conA, cell lysates were stripped of membrane-associated proteins by overnight incubation with conA Sepharose resin. (D) Western blot after 1-h treatment with Wnt-3A–conditioned media in U2OS cells transfected with the indicated siRNAs. Blots represent n = 3.
Figure 7.

Endogenous RYK and MIB1 are required for Wnt/CTNNB1 signaling at the level of receptor activation. (A) RYK and MIB1 siRNA transfection inhibits cDNA overexpression-mediated activation of BAR by Wnt-3A and Disheveled (DVL3) but not CTNNB1 or nondegradable CTNNB1* (S33A, S37A, T41A, and S45A). Data are mean + SD (error bars). (B) Western blot of cell lysates after a time course of treatment with Wnt-3A–conditioned media in cells transfected with control or MIB1 siRNAs. (C) Western blot after 1 h of treatment with Wnt-3A–conditioned media in HEK293T cells transfected with the indicated siRNAs. In rows labeled conA, cell lysates were stripped of membrane-associated proteins by overnight incubation with conA Sepharose resin. (D) Western blot after 1-h treatment with Wnt-3A–conditioned media in U2OS cells transfected with the indicated siRNAs. Blots represent n = 3.

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