Fig. 4

Taz facilitates the binding of Gli3 to PKA. A-C Co-immunoprecipitation and western analyses in wild-type C3H10T1/2 cells or gene-konckout C3H10T1/2 cells (sgTaz or sgPKA) with or without N-Shh at 100 ng/ml for 24 hrs by using Gli3, Taz or PKAc antibody. A co-IP kit from Thermo company (Pierce #26149) was used to avoid detection of the IgG heavy chain. D, E Purified recombinant GST-Taz from E. coli was incubated with PKAc or Gli3F/R protein and western analyses were performed. F-H Immunoprecipitation and western analyses in HEK293 cells expressing vector, myc-Gli3 (WT) or myc-Gli3 sextuple mutant (MT) by using myc, Taz, or PKAc antibody. I-N Immunoprecipitation and western analyses in HEK293 cells expressing myc-Taz or Taz siRNA by using PKAc, Taz or Gli3 antibody. All the experiments were either duplicated or triplicated, and the representative results were shown. The mean value was present under the bands