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. 2009 Nov 25;285(5):3319–3329. doi: 10.1074/jbc.M109.024000

FIGURE 1.

FIGURE 1.

Overexpression of the DJA proteins reduces the trafficking efficiency of hERG. A, HEK-293 (GripTite) cells were transiently transfected with 1 μg of HA-tagged hERG and 2 μg of Myc vector (vec), Myc-tagged DJA1 (A1), DJA2 (A2), or DJA4 (A4). Two days post-transfection, the cells were lysed, and protein expression was analyzed by Western blots with anti-hERG antibody for hERG visualization and anti-Myc antibody for DJA protein visualization. B and C, corresponding mean data (n = 8 experiments/condition). All of the values are normalized to the CG vector condition after background subtraction. Trafficking efficiency is calculated as FG/(CG+FG). *, p ≤ 0.01 versus vector CG. D, HEK-293 cells were transiently transfected with 1 μg of HA-tagged WT hERG or 2 μg of GFP and either Myc vector or Myc-tagged DJA1 or DJA2. Two days post-transfection the cells were analyzed as indicated in A. The membranes were blotted for hERG, GFP, tagged and endogenous DJA1 and DJA2, Hsc70, and tubulin as indicated. E, HEK-293 cells were transiently transfected with 1 μg of HA-tagged WT hERG and either Myc vector, FLAG-tagged Hsc70, or Myc-tagged DJA1 or DJA2 lacking their J domain (A1ΔJ and A2ΔJ) and analyzed as above. IB, immunoblot.