The PDZ binding motif of MARCH2 is not required for MARCH2 degradation of CFTR.

<p>(<b>A</b>) HEK293 cells were co-transfected with 3µg myc-MARCH2 or myc-ΔTPV-MARCH2 together with 3µg of HA-STX6 or several HA-CAL construct (see below), as indicated. After 48h, cell lysates were harvested and immunoprecipitated with an HA-affinity matrix. Cell lysates and immunoprecipitated materials were subjected to immunoblot analysis with the indicated antibodies. HA-CAL constructs used: HA-CAL (HA-tagged full length CAL), HA-CAL-PDZ (the HA-tagged PDZ domain of CAL) and HA-CAL-PCT (the HA-tagged PDZ and carboxyl domains of CAL) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068001#B18" target="_blank">18</a>] (<b>B</b>) HEK293 cells were co-transfected with 3µg CFTR (pCMV-CFTR) and increasing amounts of HA-ΔTPV-MARCH2 as indicated. After 48 h, cell lysates were harvested and subjected to immunoblot analysis with the indicated antibodies. Data shown are representative of at least three independent experiments.