Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2011 Mar 17;7(3):e1001335. doi: 10.1371/journal.pgen.1001335

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Molnar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PMC Copyright notice

(A–D) Effects of different Krz-Flag mutant forms expressed in the dorsal compartment on Smo expression. (A) Control ap-Gal4 UAS-GFP/+; UAS-krz-Flag/+ showing Flag (blue) and GFP (green), and the elimination of Smo (red) in the dorsal compartment. (B) ap-Gal4 UAS-GFP/+; UAS-krz^V94D-Flag/+. The mutant protein Krz^V94D is over-expressed (blue staining in B), but it does not affect Smo expression (red in B′). (C) ap-Gal4 UAS-GFP/+; UAS-krz^S427D-Flag/+. The mutant protein Krz^S427D is over-expressed (blue staining in C), and eliminates Smo expression from dorsal cells (red in C′). (D) ap-Gal4 UAS-GFP/+; UAS-krz^Leu-Flag/+. The mutant protein Krz^Leu is over-expressed (blue staining in D), but it does not affect Smo expression (red in D′). A′–D′ corresponds to the single red channels showing Smo expression. (E, F) Interactions between Krz and Smo in S2 cells. Stable Myc-Smo S2 or S2 control cells were transiently transfected with either empty pUAS/actinGAL4 vector (mock lanes), pUAS-krz/actinGAL4 (Kurtz lanes) or pAWF-krz mutant constructs (pAWF-krz^wt, pAWF-krz^V94D, pAWF-krz^S247D or pAWF-krz^Leu; F-Kurtz construct lanes) as indicated in the figure, in presence of Hh conditioned medium for 18–24 hrs. Cell extracts were immunoprecipitated with anti-Myc affinity gel. Immunoprecipitates (F) and whole-cell lysates (E) were immunoblotted with antibodies against Krz (detecting both endogenous Kurtz, lower band and the Flag-Kurtz protein, upper band), Myc (detecting Myc-Smo protein) and Tubulin. In control IPs carried out in Smo-Myc non-expressing cells transfected with pAWF-krz^wt, pAWF-krz^V94D, pAWF-krz^S247D or pAWF-krz^Leu, we did not detect any Krz or FLAG-Krz protein (data not shown). The levels of Smo were determined in the whole cellular lysates by immunoblotting. The basal amount of Smo in the Krz^wt transfected cells was defined as 1 and all data were normalised by tubulin protein levels. Data are the mean ±SEM of three independent experiments. A representative gel is shown. * (P<0.05); *** (P<0.001) *, P<0.05; ***, when compared to values of Krz^wt transfection.