Panel A shows protein schematics, illustrating human and zebrafish AQP12 constructs without hemagglutinin tags. Panel B shows immunoblot images, comparing total membrane and yolk platelet membrane localization of AQP12 proteins. Panel C shows immunofluorescence microscopy images, depicting localization of AQP12 proteins in oocytes and isolated yolk platelets. Panel D shows protein schematics, illustrating human and zebrafish AQP1 wild type and mutant constructs. Panel E shows immunoblot images, comparing membrane localization of AQP1 wild type and mutant proteins. Panel F shows immunofluorescence microscopy images, illustrating localization of human AQP1 proteins in oocytes and isolated yolk platelets. Panel G shows immunofluorescence microscopy images, depicting localization of zebrafish Aqp1aa proteins in oocytes and isolated yolk platelets.
HA epitope tag in WT AQP12 and AQP1 mutants does not affect YPM channel trafficking. (A) Schematic diagrams of HsAQP12-WT and DrAqp12-WT without the HA tag. (B) Immunoblots of TM and YPM protein extracts from X. laevis–uninjected oocytes (control) or expressing HsAQP12-WT or DrAqp12-WT probed with specific antibodies for human AQP12 and zebrafish Aqp12 (Table S1). The α-PDI antibody was used as a protein loading control. (C) Immunostaining of oocytes and YPs in control oocytes and expressing the channels. (D) Schematic diagrams of HsAQP1, DrAqp1aa, and mutant constructs used in Fig. 5, A and E lacking the HA tag. (E–G) Immunoblot (E) and immunostaining (F and G) of the corresponding channels in oocytes and YPs using a human AQP1 and a fish (Gilthead seabream, Sparus aurata) Aqp1aa-specific antibody (Table S1), and the α-PDI antibody, as indicated. Molecular mass markers (kDa) are on the left. Scale bars in C, F, and G, 10 µm (upper panels), 5 µm (lower panels). WT, wild type. Source data are available for this figure: SourceData FS3.