(A-C) Double immunolocalization of SaAqp1ab1-WT, Flag-tagged HhAqp1ab1-WT or Flag-tagged SsAqp1ab2-WT (green) with the respective HA-tagged splice forms in X. laevis oocytes as indicated. Oocytes expressing SsAqp1ab2-WT and splice forms were co-injected with halibut YwhazLb and exposed to FSK. In all panels, the plasma membrane is indicated by an arrowhead, whereas the co-localization of WT aquaporins and splice forms is indicated by arrows. Scale bars, 10 μm. (D-F) Representative immunoblots of total (TM) and plasma (PM) membrane protein extracts from oocytes injected with water (W) or co-expressing the different aquaporin WT channels with increasing amounts of the splice forms as indicated. (G-I) Illustrative immunoblots of precipitated proteins from oocytes treated as in A-C using anti-SaAqp1ab1 or anti-Flag (for HhAqp1ab1 and SsAqp1ab2) antibodies, and revealed with antibodies against SaAqp1ab1, Flag or HA. In D-I, upper blots were probed with SaAqp1ab1 (D and G) or Flag (E, F, H and I) antibodies, whereas in all panels the lower blots were probed with HA antibodies. Molecular mass markers (kDa) are on the left. The arrowhead in F and I indicate potential post-translational modifications of the SsAqp1ab1_v2 isoform.

With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S).

Peer reviewed