W287G and Y71G reduces ASIC1a glycosylation while a reduced culture temperature has an opposite effect.

<p>(<b>A & B</b>) Representative blots showing the effect of a reduced culture temperature on ASIC1a glycosylation. CHO cells were transfected with wild-type ASIC1a, W287G or Y71G mutant. Surface biotinylation was performed after culturing at 37°C (A) or 27°C (B) for 24 hrs. Lysates were treated with PNGase F or Endo H as indicated, followed by NeutrAvidin precipitation. Surface and total fraction were blotted for ASIC1a. The relative position of Endo H-resistant and -sensitive ASIC1a was indicated by arrows. Note that 27°C increased the percentage of Endo H-resistant wild-type ASIC1a in total lysate. Also note the appearance of Endo H-resistant W287G and Y71G at the cell surface at 27°C (arrows). (<b>C</b>) Quantification of Endo H-resistant wild-type ASIC1a at 37°C and 27°C. Asterisk indicates significant difference (P = 0.025); n.s. stands for not significant.