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

. 2022 Sep 30;12:16049. doi: 10.1038/s41598-022-20476-1

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

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Phenotypes of Wnt3 knockdown with siRNA in Cladonema fertilized eggs. (A) Typical morphology of Cladonema egg and planula. The wild-type Cladonema planula larvae exhibit a slight oval shape. To deliver siRNAs targeting CpWnt3, electroporation was performed in fertilized eggs (1-cell stage), and phenotypes were confirmed at planula in one day. (B, C) Phenotypes of 1-day planula after CpWnt3 siRNA knockdown. (D) Schematic of ellipse approximation for planula morphology and calculation of the aspect ratio. The aspect ratio was calculated by dividing the long axis (a) by short axis (b). (E) Boxplots showing the aspect ratio of the 1-day planula after siCpWnt3 electroporation (0 and 300 ng/µl). Center lines show the medians; x’s denote the mean values; box limits indicate the 25th and 75th percentiles; whiskers show maximum and minimum values; inner points and outliers are represented by circles. Number of examined planulae: 0 ng/µl, n = 22; 300 ng/µl, n = 50. p = 0.658303. n.s., not significant. (F) Quantification of CpWnt3 mRNA levels in 1-day planula by RT-qPCR. CpEF1alpha was used as an internal control. Bar heights represent the mean value. Error bars indicate standard deviation. CpWnt3 expression levels are standardized relative to the control (0 ng/µl). Experiments were performed in triplicate and repeated three times. p-values: 150 ng/μl, p = 0.376498; 300 ng/μl, p = 0.073956. (G, H) Representative in situ hybridization (ISH) image of oral CpBra expression in 1-day planula for control (G: 0 ng/µl siCpWnt3) and CpWnt3 siRNA (H: 300 ng/µl siCpWnt3). The oral pole is indicated by an asterisk. (I) Quantification of CpBra expression phenotypes based on ISH images. Stacked bar plots show the percentage of 1-day planulae in each phenotypic class. CpBra expression patterns are categorized into three phenotypes (strong, weak and none). 0 ng/µl, n = 74; 300 ng/µl, n = 58. Scale bars: (B, C) 200 µm, (G, H) 50 µm.