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. Author manuscript; available in PMC: 2019 Jan 1.
Published in final edited form as: Glia. 2017 Sep 14;66(1):108–125. doi: 10.1002/glia.23231

Figure 1. SUR1-TRPM4 co-immunoprecipitates with AQP4.

Figure 1

(A–E) Co-immunoprecipitation experiments in COS-7 cells transfected with AQP4 M1 or M23 isoforms, TRPM4, SUR1, or an SUR1-TRPM4 fusion protein; AQP4 immunoblots showing AQP4 M1 or M23 in total lysates (A, B lane 1), AQP4 upon TRPM4 immunoprecipitation (IP) (A, B lane 2), no AQP4 upon TRPM4 IP in cells that express AQP4 but not TRPM4 (A, B lane 3), and no AQP4 when the anti-TRPM4 IP antibody (IP AB) is omitted (A, B lane 4); TRPM4 immunoblot showing TRPM4 in total lysates (C lane 1), TRPM4 upon IP of AQP4 M1 (C lane 2) or M23 (C lane 3), no TRPM4 upon AQP4 IP in cells that express TRPM4 but lack AQP4 (C lane 4), and no TRPM4 when the anti-AQP4 IP antibody is omitted (C lanes 5 and 6); SUR1 immunoblot showing SUR1 in total lysates (D lane 1), SUR1 upon IP of AQP4 M1 (D lane 2) or M23 (D lane 3), no SUR1 upon AQP4 IP in cells that express SUR1 but lack AQP4 (D lane 4), and no SUR1 when the anti-AQP4 IP antibody is omitted (D lanes 5 and 6); SUR1 immunoblot showing the SUR1-TRPM4 fusion protein in total lysates (E lane 1), the SUR1-TRPM4 fusion protein upon IP of AQP4 M1 (E lane 2) or M23 (E lane 3), no SUR1-TRPM4 fusion protein upon AQP4 IP in cells that express SUR1-TRPM4 but lack AQP4 (E lane 4), and no SUR1-TRPM4 fusion protein when the IP antibody is omitted (E lanes 5 and 6); all results represent n=5 replicates. (F) Co-immunoprecipitation experiment in HEK 293 cells stably expressing either empty vector (−SUR1) or SUR1 (+SUR1) and transfected with AQP4 M1 and TRPM4, showing 4-fold greater TRPM4 abundance upon AQP4 IP in SUR1+ cells versus SUR1− cells (top row); TRPM4 and AQP4 in total lysates (middle, bottom rows) do not show differences in expression; n=6; *p<0.05 in t-test; densities were normalized to average SUR1− band density (dotted line).