Supplementary Figure 1: Kif7^L130P encodes a stable protein that does not localize to cilium tips.

(a) Immunoblot with Kif7 antibody in cell lysates of wild-type, Kif7^L130P and Kif7^−/− MEFs. (b) Kif7 (green) is absent from Kif7^L130P cilia regardless of pathway activation. Acetylated α-tubulin (red) stains cilia and γ-tubulin (blue) stains basal body. Arrows indicate Kif7 at the transition zone. Scale bar, 1 μm. (c) Kif7 localizes to cilium tip in Smo^−/− and (d) Gli2^−/−Gli3^−/− double mutant MEFs. Acetylated α-tubulin (red) stains cilia and γ-tubulin (blue) in (c) and magenta in (d) stains basal body. Scale bar, 5 μm. (e) GM130 (green), a Golgi membrane protein, appears normal in Kif7^L130P MEFs. DAPI in blue. Scale bar, 17 μm. (f) In contrast to Kif7 knockdown cells¹⁴, normal numbers of centrosomes and cilia are present in wild-type and Kif7^L130P MEFs. Acetylated α-tubulin (red) marks axonemal microtubules and γ-tubulin (magenta) marks basal bodies. DAPI is in blue. n = 165 cilia were analysed and were pooled from 4 independent experiments. Error bars represent the s.d., P > 0.0001 by student t-test. Scale bar, 22 μm.