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Figure 7. Overexpression of Spop rescues phenotypes caused by partial knockdown of Dzip1. (A) Western blot results showing that increases to Gli stability following Dzip1 depletion were reversed by Spop overexpression. RNAs encoding MT-Gli1-3 (2 ng) and MT-GFP (50 pg) were injected, alone, or in combination with MT-Spop (100 pg) into controls or embryos that had received a prior injection of DMO. Embryo were harvested at the neurula stage and subjected to Western blot with the anti-myc antibody to detect the levels of Spop. (B) Forced Spop expression decreases the stability of xGli1, xGli2, and hGli3 in NIH3T3 cells. NIH3T3 cells were transfected with MT-Glis and MT-GFP, alone, or in combination with increasing levels of MT-Spop. Western blots were carried out with the cell lysates to determine the levels of Gli1, 2 and 3. GFP serves as a transfection and loading control. (C) In situ hybridization results showing the expression of ptc1, pax6, and pax2 in controls or embryos injected with DMO alone or DMO and Spop RNA. �inj� indicated the side which received Spop RNA injection. (D-F) Summary of the ptc1 (D), pax6 (E), and pax2 (F) expression phenotypes in controls and injected embryos. |
