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. 2019 Sep 4;132(17):jcs230680. doi: 10.1242/jcs.230680

Fig. 4.

Fig. 4.

Lack of Ahi1 leads to reduced Shh signaling activation but unaltered ciliary trafficking of Smo. (A) Serum-starved Ahi1+/+ and Ahi1−/− MEFs (24 h) were treated with DMSO (control) or SAG (a Shh activator) for 24 h. Cells were co-immunolabeled for Smo (red) and acetylated α-tubulin (Ac-tub; green) with DNA/nuclei (blue). Smo localization is only seen in cells treated with SAG (arrows). Scale bar: 5 µm. (B) Graph showing Smo intensities at primary cilia after Shh activation by SAG. n>100 cilia per group (n=4/genotype). Relative expression of Gli1 (C) and Ptch1 (D) mRNAs in Ahi1+/+ and Ahi1−/− MEFs after DMSO (control) or SAG treatment for 24 h. Fold change was normalized to Ahi1+/+ cells (set at 1) treated with DMSO after normalization (Rpl13a mRNA). Bars represent means from n≥3 cell lines/genotype and experiments were carried out in duplicate. Error bars represent s.e.m. ****P<0.0001; **P<0.01; *P<0.05; ns, not significant (Mann–Whitney test for B and unpaired two-tailed t-tests for C and D).