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. 2008 Sep;22(9):3255–3263. doi: 10.1096/fj.07-105338

Figure 3.

Figure 3.

Preferential interaction of Aha1 with ΔF508 CFTR is reduced by ΔFRD. A) HEK293 cells were transfected as follows: wt CFTR plus EGFP (lane 2); wt CFTR plus ΔFRD (lane 3); ΔF508 CFTR plus EGFP (lane 4); ΔF508 CFTR plus ΔFRD (lanes 5 and 6). Equal amounts of total protein (∼50 μg) were subjected to IP with anti-CFTR (lanes 2–5) or anti-V5 (lane 6) monoclonal antibodies. Of the total protein used in the IP of lane 2, 5% was loaded in lane 1 as input. Coprecipitated Hsp70 (top panel), Hsp90 (middle panel), or precipitated CFTR (bottom panel) were detected by IB. HC, antibody heavy chain. B) HEK293 cells were transfected with ΔF508 CFTR plus EGFP (lanes 1, 7); ΔF508 CFTR plus ΔFRD (lanes 2–4); wt CFTR plus EGFP (lane 5); wt CFTR plus ΔFRD (lanes 6). Wt or ΔF508 CFTR:ΔFRD plasmid DNA ratios are indicated above the panels. Following co-IP (protocol identical to A), IBs were performed with anti-Aha1 or anti-CFTR. Relative Aha1 density (given below panel) gives the average of two independent experiments. Total proteins (30 μg) were loaded for detection of ΔFRD or Aha1 expression (bottom panels).