Xenbase Image ID: 129960

Fig. 1. Growth characteristics of the Xenopus proximal tubules. (A–A′′) Representative immunofluorescence analysis of proximal tubules at stage 42 marked by 3G8 (green); mitotic cells were labeled by pH3 (red), and nuclei were visualized by DAPI (blue). Asterisks indicate dividing cells in the proximal tubules. (B and C) Graphs depicting the total number of proximal tubular cells (B) and the percentage of pH3-positive cells (C) in uninjected controls and xDicer-MO–injected embryos at stages 37 and 42. The number of embryos analyzed is indicated in the bars. *P < 0.005; ns, not statistical significant change. (D and D′) TUNEL staining (red) of stage 42 embryos; images were counterstained with DAPI (blue) and ECL (green) to visualize nuclei and proximal tubules, respectively. Inset in D shows a DNase I-treated sample as positive control. (E) Total number of cells present in the proximal and distal tubule of a single pronephros during stages 34–46 [equivalent of 45–106 h postfertilization (hpf)]. Proximal tubular growth follows a sigmoidal trend (blue line, with an R2 = 0.9274), whereas the distal tubular one is linear (black line, with an R2 = 0.6914). Inset shows scheme subdividing proximal tubular expansion into three distinct phases, baseline (B, stages 34–37), growth (G, stages 38–42), and stationary (S, stages 43–46). The number of embryos analyzed is presented in Dataset S1. (F) Scatterplot depicting total number of cells and those in mitosis; each individual embryo used for the time course analysis in E from stages 34–46 is represented by a blue diamond. Note that the number of mitotic cells does not directly correlate with the overall cell numbers, but instead follows a polynomial trend (red line, with an R2 = 0.8009). Black line indicates an extrapolation of a direct correlation considering only proximal tubules of 500 cells or fewer (with an R2 = 0.7604). (G–U) Immunofluorescence staining of sectioned proximal tubules at different developmental stages using antibodies recognizing pErk1/2[T202/Y204], pAkt[Ser473], and pS6[Ser235/236]. Tubular structures are outlined by yellow dotted lines. Inset in I shows pErk1/2 staining in the somites as control for antibody activity. Insets in P and U show immunofluorescence analysis of total Akt and ribosomal S6, respectively. All error bars correspond to the SD. Image published in: Romaker D et al. (2014) Copyright © 2014. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Experiment + Assay Source Phenotypes and Disease Xla Wt + dicer1 MO + NF37/38-42 (immunohistochemistry) fig.1.b Xla Wt + dicer1 MO + NF42 (immunohistochemistry) fig.1.b Anatomical Phenotype abnormally decreased number of proximal tubule Xla Wt + dicer1 MO + NF37/38-42 (immunohistochemistry) fig.1.c Anatomical Phenotype decreased cell population proliferation in proximal tubule Larger Image Printer Friendly View

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