A Hh- and SAG-responsive NIH 3T3/Smo-mEos2 cell line.

<p>(A–C) The NIH 3T3/Smo-mEos2 cells were incubated in the absence or presence of SAG or ShhN, and then fixed and stained with anti-acetylated tubulin (primary cilium, red) and DAPI (nucleus, blue). Smo-mEos2 was detected in the primary cilium only when the cells were treated with 100 nM SAG (B) or ShhN (C), but not in the un-treated cells (A). The boxed region in each main panel was viewed on the right in separated red, green channels, and shifted overlays. Scale bar, 5 µm. (D) Mean intensity of Smo-mEos2 fluorescence in cilia of NIH 3T3/Smo-mEos2 cells treated with ShhN or SAG. Each point shows the mean ± SD of fluorescence from 10–20 cilia. (E) The NIH 3T3/Smo-mEos2 cells were incubated in the absence or presence of ShhN or SAG. The mRNA was extracted 48 hours later and real-time quantitative RT-PCR was performed to measure the mRNA level of Gli1.