FIGURE 8.

Subcellular fractionation experiments performed to determine the effect of the Y1424A mutation on CFTR recruitment CCV in CFBE41o- cells. The CCV fractions were prepared by density gradient and differential centrifugation in parental CFBE41o- cells 72 h after transfection with the GFP-CFTR WT or GFP-CFTR Y1424A mutant. A, a representative Western blot (IB) demonstrates successful isolation of CCVs as demonstrated by the enrichment of clathrin in the CCV fractions. P2, cytosolic fraction; S2, microsomal fraction. A representative Western blot (B) and summary of experiments (C) demonstrate that the Y1424A mutation did not decrease the abundance of CFTR in CCVs. The CCV abundance of CFTR was normalized for the abundance of CFTR in PNS. Three experiments/group were performed. Error bars, S.E.