Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Jul 2;217(7):2485–2501. doi: 10.1083/jcb.201710019

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2018 Dionne et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 5. — CA causes cystogenesis. (A) H&E-stained kidney sections from postnatal (P15) Hoxb7-Plk4, Six2-Plk4, and control mice. (B) Immunostaining of P15 and P7 sections for centrosomes (γ-tubulin), cilia (acetylated α-tubulin), proximal tubules (LTL), collecting ducts (DBA), and nuclei (DAPI). Cells from control animals contained the normal complement of centrosomes and assembled a single cilium. In contrast, CA disrupted ciliogenesis, resulting in aciliated cells or formation of short cilia. White box denotes regions shown in insets. Arrowheads point to cells with multiple centrosomes. Bars (insets), 2 µm. (C) Graph illustrating a decrease in the fraction of ciliated cells in collecting ducts of P0 to P15 Hoxb7-Plk4 kidneys. n = 1,971 cells from 186 tubules (P0 WT), 1,769 cells from 135 tubules (P0 Hoxb7-Plk4), 2,550 cells from 411 tubules (P15 WT), and 3,541 cells from 149 cysts (P15 Hoxb7-Plk4). (D) Quantification of the length of cilia in cystic epithelia of Hoxb7-Plk4 kidneys, demonstrating a small but significant decrease in mean ciliary length by P15. n = 951 cilia measured (P0 WT), 314 cilia (P0 Hoxb7-Plk4), 1,852 cilia (P15 WT), and 1,186 cilia (P15 Hoxb7-Plk4). (E) Graph demonstrating a decrease in the fraction of ciliated cells in proximal tubules of P0 to P7 Six2-Plk4 kidneys. n = 2,218 cells from 251 tubules (P0 WT), 1,637 cells from 188 tubules (P0 Six2-Plk4), 643 cells from 91 tubules (P7 WT), and 678 cells from 24 cysts (P7 Six2-Plk4). (F) Graph illustrating decreased ciliary length in cystic epithelia of Six2-Plk4 kidneys by P7. n = 1,363 cilia measured (P0 WT), 811 cilia (P0 Six2-Plk4), 329 cilia (P7 WT), and 216 cilia (P7 Six2-Plk4). For all graphs, a two-tailed unpaired t test was performed to determine statistically significant differences (*, P < 0.05). (G and H) Representative immunoblots of kidney lysates demonstrating that phosphorylated ribosomal protein S6 (p-S6) and p-S6K increase upon CA. Two examples are shown for control (WT) and Hoxb7-Plk4. Kidney lysates obtained from Hoxb7-PKD1^f/f mice were used as a positive control for pathway activation. Total levels of S6 were mostly unchanged, whereas total S6K was also elevated. Graph shows quantifications of band signal intensity, normalized to actin for each marker. n = 3 kidneys from each genetic sample; (*, P < 0.05). (I) Representative images of P15 kidney sections from control and Hoxb7-Plk4 mice, immunostained with antibodies against Ki-67, DBA, and DAPI. Proliferating Ki-67–positive cells are highlighted with white arrowheads. (J) Graph denoting the percentage of Ki-67–positive cells in DBA-positive collecting ducts (WT) or cysts (Hoxb7-Plk4). A two-tailed unpaired t test was used to determine statistically significance (*, P < 0.05).