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. 2009 Dec 14;4(12):e8284. doi: 10.1371/journal.pone.0008284

Figure 2. N-cadherin reduces cell proliferation via interaction with LRP5 and Wnt signalling.

Figure 2

(A) Immunoprecipitation analysis showing N-cadherin and LRP5 interaction in control (Flag) cells and N-cadherin (N-Cad) overexpressing cells which is blocked by N-cadherin antibody. Cells were treated with blocking N-cadherin antibody (Ab) or control antibody (IgG) for 24 hours, cell lysates were immunoprecipitated (IP) with N-cadherin antibody and analysed by Western-blot (WB) with LRP5 antibody. LRP5, N-cadherin and GAPDH in total proteins were used as loading controls. (B) Treatment with Wnt3a or blocking N-cadherin antibody restores cell proliferation in N-Cad cells. Flag and N-Cad cells were transfected with DKK1 expression vector or empty vector (EV), treated with Wnt3a CM (15%) or N-cadherin antibody for 24 hours and cell replication was determined. Means are +/− SD. Values that are significantly different are indicated (a, P<0.05 vs untreated Flag EV cells; b, P<0.05 vs Flag EV cells treated with N-Cad Ab or Wnt3a; c, P<0.05 vs N-Cad EV cells treated with N-Cad Ab or Wnt3a). (C) N-cadherin silencing increases osteoblast proliferation. Flag cells were transfected with a specific N-cadherin si-RNA or a non relevant si-RNA (si-NR) and treated with Wnt3a CM (15%) for 24 hours and cell replication was determined (a, P<0.05 vs -Wnt si-NR treated cells; b, P<0.05 vs -Wnt si-N-Cad treated cells).