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. Author manuscript; available in PMC: 2015 Feb 19.
Published in final edited form as: Biochem J. 2012 May 1;443(3):789–798. doi: 10.1042/BJ20112062

Figure 3. Wnt5a is required for Ihh-induced Nkx3.2 degradation.

Figure 3

(A) Nkx3.2-HA or HA-Cbfβ expression construct was transfected into ATDC5 cells in the absence or presence of Wnt5a expression plasmid for 48 h. Total cell lysates were analyzed by western blotting.

(B) Expression vehicles for control or Wnt5a shRNA were transfected into ATDC5 cells with Nkx3.2-HA expression construct in the absence or presence of SmoA1-HA expression plasmid for 48 h. Total cell lysates were analyzed by western blotting.

(C) ATDC5 cells were infected with control or Wnt5a shRNA-harboring lentivirus. At 24 h post-infection, cells were treated with DMSO or 10 μM PMP. After an additional 24 h, total cell lysates were analyzed by western blotting.

(D) Wnt5a expression plasmid was transfected into ATDC5 cells in the absence or presence of control or Wnt5a shRNA constructs, and total cell lysates were analyzed by western blotting after 48 h.

(E) ATDC5 cells were transfected with Nkx3.2-Myc or Wnt5a expression plasmids. At 24 h post-transfection, cells were treated with 0.5% DMSO or 5 μM CMP for an additional 24 h, and total cell lysates were analyzed by western blotting.