Figure 4. .
GLUK6Δ present on the plasma membrane of transfected HEK293 cells. a, Membrane proteins biotinylated on HEK293 cells overexpressing GLUK6 wild type (WT) or GLUK6Δ (Delta) at 48 h after transfection (Pinpoint [Pierce]). Nontransfected HEK293 cells served as a control (Mock). After cell lysis (Total Lysate samples), biotinylated membrane proteins were bound to a NeutrAvidin affinity column. Nonbiotinylated protein was separated, because it does not bind to the NeutrAvidin affinity column (Flow Through samples). Membrane proteins were eluted with sample buffer (Eluate samples). All fractions were separated on a 10% SDS gel, were blotted, and were probed for GLUK6 (with 05-921 [Upstate Biotechnology] at 1:800). GLUK6 WT and GLUK6Δ are present in the membrane protein fraction (Eluate samples). b, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) loading control (ab9482 [Abcam]).