FIGURE 2.

Establishing a culture system to investigate HCN2 channel SUMOylation. Cell lysates from a Hek cell line stably expressing GFP-HCN2 channels (A) and the parental Hek cell line (B) were used in IP experiments with an antibody against GFP. WBs containing the lysate (L) and IP products (IP) were then probed with anti-HCN2 and anti-GFP antibodies. A doublet was recognized by both antibodies in the stably transfected but not parental Hek cell line, suggesting that the doublet represents GFP-HCN2 channels. The band present at 50 kDa in the IP lanes corresponds to the heavy chain of the anti-rabbit antibody used in the IP experiment. The anti-rabbit secondary antibody used in the GFP WB produced a strong signal. The anti-goat secondary antibody used in the HCN2 WB produced a much weaker signal.