Fig. S3.

CCDC22 mutants display reduced binding to VPS33B/GST-VIPAR in vitro. (A) Myc-tagged CCDC22 was produced by in vitro transcription/translation and then subjected to GST pull-down (PD) using VPS33B/GST-VIPAR or GST alone. Samples were analyzed by immunoblotting with anti-myc. Point mutations of CCDC22 demonstrate reduced binding to VPS33B/GST-VIPAR compared to the wild-type (wt) protein. (B) Quantitation of relative binding efficiency across three independent experiments. Immunoblot band intensities were quantified and normalized (intensity of PD band divided by input band, normalized to the PD efficiency of the wild-type construct in each experiment).