Figure 3. Wnt-11 stabilized cell surface... | Rockefeller University Press

Figure 3.

$Figure 3. Wnt-11 stabilized cell surface PAPC, whereas C-cadherin remained unaffected. (A) Cell surface biotinylation assay performed with DMZ explants. Cell surface and total protein amount of xPAPC-myc (9E10) and endogenous xC-cadherin (6B6) were detected by Western blotting. xWnt-11 depletion reduced cell surface amount of xPAPC-myc, whereas cell surface level of endogenous xC-cadherin was not effected. neg. KO, negative control. (B) Relative cell surface amounts of xPAPC-myc and endogenous xC-cadherin in dependency of xWnt-11, calculated as described in Materials and methods. Error bars show SEM. Student’s t test was performed (*, P < 0.05). # exp., number of independent experiments. (C) Biotin pulse-chase experiment performed with DMZ explants. At the indicated induction times, the residual biotin-labeled (surface) and total protein amount of xPAPC-myc (9E10) and endogenous xC-cadherin (α-Ccad) were detected by Western blotting. Relative surface amounts of xPAPC-myc and endogenous xC-cadherin were plotted over endocytosis induction time. Cell surface xPAPC was stabilized in xWnt-11–overexpressing DMZ explants, whereas xWnt-11 depletion led to a faster degradation of xPAPC membrane fraction. xWnt-11 did not affect the cell surface expression of xC-cadherin. Injection amount was as follows: 1 ng xPAPC-myc RNA, 40 pg xWnt-11 RNA, and 1 pmol xWnt-11 MO. pos. KO, positive control. n, number of independent experiments.$

Wnt-11 stabilized cell surface PAPC, whereas C-cadherin remained unaffected. (A) Cell surface biotinylation assay performed with DMZ explants. Cell surface and total protein amount of xPAPC-myc (9E10) and endogenous xC-cadherin (6B6) were detected by Western blotting. xWnt-11 depletion reduced cell surface amount of xPAPC-myc, whereas cell surface level of endogenous xC-cadherin was not effected. neg. KO, negative control. (B) Relative cell surface amounts of xPAPC-myc and endogenous xC-cadherin in dependency of xWnt-11, calculated as described in Materials and methods. Error bars show SEM. Student’s t test was performed (*, P < 0.05). # exp., number of independent experiments. (C) Biotin pulse-chase experiment performed with DMZ explants. At the indicated induction times, the residual biotin-labeled (surface) and total protein amount of xPAPC-myc (9E10) and endogenous xC-cadherin (α-Ccad) were detected by Western blotting. Relative surface amounts of xPAPC-myc and endogenous xC-cadherin were plotted over endocytosis induction time. Cell surface xPAPC was stabilized in xWnt-11–overexpressing DMZ explants, whereas xWnt-11 depletion led to a faster degradation of xPAPC membrane fraction. xWnt-11 did not affect the cell surface expression of xC-cadherin. Injection amount was as follows: 1 ng xPAPC-myc RNA, 40 pg xWnt-11 RNA, and 1 pmol xWnt-11 MO. pos. KO, positive control. n, number of independent experiments.