Supplementary Figure 10: Validation of the 7e12 antibody to PC1. | Nature Genetics

Supplementary Figure 10: Validation of the 7e12 antibody to PC1.

From: Mutations in DZIP1L, which encodes a ciliary-transition-zone protein, cause autosomal recessive polycystic kidney disease

Supplementary Figure 10

(a,b) PC1 antibody (mAb 7e12 raised to the N-terminal leucine rich repeat region of PC1 (24-180aa), aliquots obtained from both Abcam and Santa Cruz Biotechnology – see online methods for details) detected overexpressed PC1 protein (green) in IMCD3 cells. (c-j) PC1 staining (green) on the ciliary membrane in Pkd1 or Pkd2 heterozygote control cells, and lack of staining in null mutant cells, with the PC1 7e12 antibody preparation indicated. Acetylated-α-tubulin (magenta) marks the ciliary axoneme. (k,l) Quantification of the percentage of ciliated cells with PC1 along the axoneme in Pkd1 or Pkd2 heterozygous control cells. No homozygous mutant cells analysed displayed detectable PC1 staining along the axoneme. Note that PC2 is required for ciliary trafficking of PC1, so axonemal PC1 staining would not be expected in Pkd2 null cells. Cilia were scored from two independent experiments. Approximately 100 ciliated cells were analysed in each experiment. The values are not compared statistically. Error bars, s.e.m. Scale bars = 1μm. See online methods for details of the Pkd1 and Pkd2 cell lines used for these validation experiments.

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