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. 2010 Aug;24(8):3103–3112. doi: 10.1096/fj.09-141788

Figure 5.

Figure 5

Maturation of coexpressed CFTR halves containing ΔF508 is not rescued by suppressor mutations. N and C halves of CFTR were coexpressed in HEK293 cells. N halves were either WT or carried the ΔF508 mutation, in the absence or presence of suppressor mutations (3S: G550E/R553M/R555K). The two halves were split either between NBD1 and the R domain (1–634+634–1480) (A), or between the R domain and MSD2 (1–830+837–1480) (B). At 48 h after transfection, cells were solubilized, and lysates were immunoprecipitated with L12B4 conjugated to protein A agarose beads. Bound proteins were eluted with 2× SDS-PAGE sample buffer, and eluates were reduced and subjected to Western blot analysis using anti-CFTR antibodies mAb13-4 and mAb596, to detect the N- and C-terminal halves, respectively. Goat anti-mouse IgG1-IR800 and IgG2b-Alexa680 were used to detect both halves of CFTR simultaneously in the same blot. Arrows with asterisks indicate complex glycosylated forms of the C halves (n=3).