Fig. 2

BCA assay and Western Blot analysis was conducted on supernatant fractions containing all membrane proteins extracted with DIBMA, Carboxy-DIBMA, DDDG, DDM, l LMNG or FOS-Choline (FOSC) from adherent HEK293 (blue), K. phaffii (orange), and Expi293F suspension cells (purple). A-E) Yields of extracted membrane proteins for DIBMA and Carboxy-DIBMA at different polymer/membrane mass ratios: R = 0.25 (A), R = 0.5 (B), R = 0.75 (C), and R = 1.0 (D). E) Yields of extracted membrane proteins for DDDG, DDM, LMNG and FOS-Choline (FOSC). Except FOS-choline, which resulted in higher protein yields, and membrane-protein extraction from K. phaffii, all extraction agents yielded similar protein yields (50–60%) from all three expression systems. The membrane-protein concentration was normalized to the total membrane-protein concentration before solubilization and expressed as a percentage yield (%). F-G) Western Blot analysis was conducted on the supernatant fractions extracted from adherent K. phaffii (F), adherent HEK293 cells (G) and Expi293F (H) cell membranes using DIBMA and Carboxy-DIBMA at various polymer-to-membrane ratios ranging from R = 0.25 to 1.0, as well as DDM, DDDG, FOS-Choline (FOSC), and LMNG. A specific TRPC3 antibody was used to target TRPC3. F) DIBMA at R = 1.0 and DDM extract the highest quantity of TRPC3. G) DIBMA extracts the highest quantity of TRPC3. H) DDM and DDDG extract the highest amount of TRPC3. Note: Please note that the Western blots on the right side of Panels F and H were cropped from the same blot. For the full, uncropped Western blot, refer to Supplementary Information, Figure S6.