Fig. 3.

L-WNK1 increases whole cell BK expression in HEK293 L-WNK1 knockout (KO) cells. A: lack of endogenous L-WNK1 expression in HEK293 L-WNK1 KO cells. Lysates obtained from HEK293 and HEK293 L-WNK1 KO cells were subjected to immunoblot with an anti-L-WNK1 antibody as described in materials and methods. Each lane was loaded with 12 μg of protein. B: L-WNK1 increases whole cell BK expression in HEK293 L-WNK1 KO cells. Whole cell lysates from HEK293 L-WNK1 KO cells transfected with GFP (control), L-WNK1, BK α-subunit, or BK α-subunit with L-WNK1 were subjected to immunoblot with an anti-HA antibody to detect L-WNK1, an anti-myc antibody to detect BK α-subunit, or an anti-actin antibody. Lanes were loaded with 1–6 μg of protein. Representative of 4 independent experiments. C: linearity of the Western blot detection system for BK α-subunit abundance. The relative intensity of the bands in the immunoblot shown in B was calculated with ImageJ. Data from cells transfected with BK α-subunit alone are shown as open symbols while those from cells transfected with the BK α-subunit and L-WNK1 are shown as gray symbols. Data were analyzed by linear regression.