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. 2015 Nov 24;10:5. doi: 10.5334/1750-2187-10-6

Figure 1.

Figure 1

Frizzled-4 Mediates Norrin induced Lef/Tcf-dependent transcriptional activation. (A) Increasing concentrations of mouse Frizzled-4 (mFz4) were cotransfected into HEK293 cells with 1 ng of human hLRP5 construct (hLRP5) and stimulated with 200 ng/mL of recombinant Norrin and the Lef/Tcf-dependent luciferase enzyme induction was quantified as described in Materials and Methods with the exception that the basal values were determined for each condition in duplicate. (B) Increasing concentrations of hLRP5 coreceptor were cotransfected into HEK293 cells with 2 ng of mFz4 and stimulated with 200 ng/mL of recombinant Norrin and the Lef/Tcf-dependent luciferase enzyme induction was quantified with the exception that the basal values were determined for each condition in duplicate. (C) HEK293 cells cotransfected with 0.25 ng of mFz4 and 1 ng hLRP5 were stimulated with increasing concentrations of recombinant Norrin and the Lef/Tcf-dependent luciferase enzyme induction was quantified. (D) Transiently transfected V5-tagged mFz4 (V5mFz4) in HeLa cells were detected by means of indirect immunofluorescence (IIF) in the absence of or following a cell permeabilization step using a V5 primary antibody.