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. 2018 Jul;94(1):665–673. doi: 10.1124/mol.117.110510

Fig. 2.

Fig. 2.

Treatment with PGRMC1 ligand AG205 decreases IR plasma membrane levels. Western blot analysis of plasma membrane (PM) protein levels in A549 (A) and HUH7 (B) cell lines from control (lanes 1 and 2) and AG205 (20 μM) treated (lanes 3 and 4, respectively) labeled with biotin and purified by avidin agarose. Proteins that were not detected at the plasma membrane are indicative of cytoplasmic (cyto) proteins (lanes 1 and 2). The Novus insulin receptor β antibody (NBP2-12793, mapping to the region between residues 1332 and 1382) was used to detect IR levels. Proliferating cell nuclear antigen serves as a control for intracellular proteins. Coomassie Blue–stained gel represents total protein levels in A549 (C) and HUH7 (D) cell lines, respectively. (E and F) Western blot analysis of total IR protein levels in A549 cells and HUH7 cells ± AG205 treatment. IR plasma membrane levels were significantly reduced after treatment with AG205 in three independent replicates of both A549 (G) and HUH7 (H) cells (P < 0.001; P < 0.01, respectively). **P < 0.01; ***P < 0.001 compared with the vehicle-treated group.