FIG 2.

Shh induces Sufu-Gli1 moving into and Sufu-Gli3 out of the nucleus. (A and B) Gli2^−/−; Gli3^−/− double mutant MEFs were treated with control 293T-CM or ShhN-CM and analyzed by IHC for anti-Gli1 (A) or anti-Sufu (B). Quantification for panels A and B was done in duplicate samples and is shown in panel C (n ≥ 35). (D) Gli2^−/−; Gli3^−/− MEFs carrying a genomically integrated shGli1-expressing unit (Gli-null) were treated with control 293T-CM or ShhN-CM and analyzed by IHC for anti-Sufu. Quantification was done in duplicates and is shown in panel E (n ≥ 38). (E) Quantification of results in panel D. (F) Western analysis of Gli1 expression in Gli1^−/−; Gli2^−/− (shNS) and Gli-null (shGli1) MEFs showing the knockdown effect of shGli1. (G and H) Anti-Gli3 (G) and anti-Sufu (H) IHC staining in Gli1^−/−; Gli2^−/− double mutant MEFs. The cells were treated with control 293T-CM or ShhN-CM for 24 h before the analysis. (I) Quantification of results in panels G and H (n ≥ 50). (J and K) Representative immunofluorescence images (J) and subcellular distribution (K) of Sufu-GFP transiently expressed in normal MEFs. (L to O) Kinetics of nuclear accumulation of Sufu-GFP in wild-type (L), Gli1^−/− (M), Gli2^−/−; Gli3^−/− (N), and Gli-null (O) MEFs.