Supplementary Figure 11: Validation of antibody to PC2 and localization of SMO in Dzip1l^wpy/wpy MEFs.

(a, b) PC2 staining (green) on the ciliary membrane in Pkd2 heterozygote control but not null mutant cells with the PC2 antibody (MV12). Acetylated-α-tubulin (magenta) marks the ciliary axoneme. (c) Quantification of the percentage of ciliated cells with PC2 along the axoneme in Pkd2 heterozygote and homozygote mutant cells. Statistical analysis based on an unpaired two-tailed Student’s t-test, t(5)=2.57,**p=0.0033. Cilia were scored in three experiments, with cells from two independent coverslips analysed for each experiment (approximately 100 ciliated cells scored on each coverslip). (d,e) In response to treatment with the HH signalling agonist SAG, SMO (green) localizes along the axoneme (acetylated-α-tubulin, magenta) in both Dzip1l^+/+ and Dzip1l^wpy/wpy MEFs. (f) quantification shows no difference in SMO localization between the two genotypes (n=4 MEF cell lines derived from individual embryos, analysed across two experiments; 28-91 cilia scored per cell line, total 299 Dzip1l^+/+ cilia and 137 Dzip1l^wpy/wpy cilia scored). Statistical analysis based on an unpaired two-tailed Student’s t-test, t(6)=0.0292, p=0.9776, ns, not significant. Error bars, s.e.m. Scale bars in a,b = 1μm; d,e = 2μm.