Native imagequant software

Pulse-chase analyses were performed using metabolic radiolabelling with [³⁵S]methionine/cysteine (Perkin Elmer, Waltham, MA, USA) and subsequent immunoprecipitation of specific radiolabeled proteins were conducted as previously described (Alexander, Schwartz, Balber, & Bangs, 2002 (link); Silverman et al., 2013 (link)). Pulse times: 2 min for VSG, 10 min for TbCatL and 15 min for p67. Chase times are specified in the relevant figures. Immunoprecipitated proteins were analyzed by SDS-PAGE followed by phosphorimaging of dried gels using a Molecular Dynamics Typhoon system with native ImageQuant Software (GE Healthcare, Piscataway, NJ).

TbVps41 RNAi trypanosomes were either untreated or treated with 1 μg/ml tetracycline and used for pulse chase analysis. Pulse/chase analyses were performed using metabolic radiolabeling with [³⁵S]methionine/cysteine (Perkin Elmer) and subsequent immunoprecipitation of specific radiolabeled proteins from cell lysates as described previously (35 (link), 36 (link)). For p67, pulse time was 15 min and for TbCATL pulse time was 10 min. Chase times are as indicated in the relevant figures. Immunoprecipitated proteins were analyzed by SDS-PAGE and phosphorimaging using a Typhoon FLA 9000 with native ImageQuant Software (GE Healthcare).