Journal: PLoS ONE
Article Title: Requirement for a Uroplakin 3a-Like Protein in the Development of Zebrafish Pronephric Tubule Epithelial Cell Function, Morphogenesis, and Polarity
Figure Lengend Snippet: Expression of upk3l in the zebrafish pronephros. (A) Dendrogram of the UP3 family (TreeFam accession TF336628 ) Legend: Bt, Bos taurus ; Cf, Canis familiaris ; Dr: Danio rerio ; Gg, Gallus gallus ; Hs, Homo sapiens ; Md, Monodelphis domestica ; Mm, Mus musculus ; Mu, Macaca mulatta ; Pt, Pan troglodytes ; Rn, Rattus norvegicus ; Xt, Xenopus tropicalis . Upk3l (A2CE76) and human (hsUP3a) are highlighted. (B) TMHMM plot and predicted structure of Upk3l: CT, cytosolic tail; EL, extracellular loop; TM, transmembrane domain. (C) Expression of upk3l message in 1-dpf and 2-dpf embryos as detected by RT-PCR; control: reaction in which the RNA template was substituted with water. (D) Western blot analysis of Upk3l expression in uninjected embryos or those injected with upk3l -MO. Acetylated-tubulin (Ac-tub) was used as a loading control. (E) Localization of Upk3l, Na + /K + -ATPase, and nuclei in cross sections of 2-dpf embryos. The Upk3l primary antibody was added to the samples in the upper row of images, but excluded from those in the lower set. The approximate location of the sections is shown in the cartoon above. Legend: G, glomerulus; N, neck; PCT, proximal convoluted tubule; PST, proximal straight tubule; DE, distal early tubule; DL, distal late tubule; CD, collecting duct. (F) Distribution of Upk3l, Na + /K + -ATPase, and nuclei in the PTs of 2-dpf embryos incubated with control MO (upper panels) or upk3l -MO (lower panels). The lumen is marked with an asterisk. (G) Immunolabeling of MDCK cells transiently transfected with cDNA encoding upk3l , human (h)UP3a alone, or hUP3a and hUP1b. Samples were costained with TRITC-phalloidin to label the cortical actin cytoskeleton and/or TO-PRO-3 to label the nuclei. Images are XZ confocal sections.
Article Snippet: The α6F monoclonal antibody supernatant to the Na+ /K+ -ATPase was obtained from the Developmental Studies Hybridoma Bank (Iowa City, IA) and used at a 1∶50 dilution.
Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Injection, Incubation, Immunolabeling, Transfection