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

letter

. 2024 Apr 15;15(9):704–709. doi: 10.1093/procel/pwae018

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

© The Author(s) 2024. Published by Oxford University Press on behalf of Higher Education Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 2. — CCDC188 cooperates with SUN5 and PMFBP1 and mediates their interaction. (A) Immunoblotting of PMFBP1 and SUN5 was performed in the testis protein lysates of WT mice and Ccdc188-KO mice. (B) Relative protein levels of PMFBP1 and SUN5 as analysed of blots in A. Data are presented as the mean ± SD; Student’s t test; ns, not significant. (C and D) Immunofluorescence staining of SUN5 (C) and PMFBP1 (D) on sperm from WT mice and Ccdc188-KO mice. Flagella and nucleus were stained with ac-TUB and DAPI, respectively. Scale bar, 5 μm. (E and F) Immunofluorescence staining of SUN5 (E) and PMFBP1 (F) on sperm from the proband and a normal control. Scale bar, 5 μm. Arrowheads in (C–F) indicated the signals of SUN5 or PMFBP1. (G) Testicular interaction components of CCDC188 were identified by IP-MS (using anti-CCDC188 antibody or anti-IgG antibody). SUN5 and PMFBP1 were identified as potential CCDC188-interacting proteins and all candidates were listed in Table S3. (H) Flag-tagged CCDC188 could be immunoprecipitated by Myc-tagged SUN5 in HEK293T cell extracts. (I) Myc-tagged PMFBP1 was immunoprecipitated by Flag-tagged CCDC188 in HEK293T cells. (J) Flag-tagged CCDC188 was transfected into HEK293T cells with Flag-tagged PMFBP1 and Myc-tagged (or Flag-tagged CCDC188 alone). Anti-Myc antibody was utilized to immunoprecipitate Myc-tagged SUN5. Flag-tagged PMFBP1 was only immunoprecipitated by Myc-tagged SUN5 in HEK293T cells when Flag-tagged CCDC188 was co-transfected. (K) Schematic diagram illustrating the role of CCDC188 in sperm head and tail connection. CCDC188 connects SUN5 and PMFBP1. Knockout of Ccdc188 in mice or loss-of-function mutation of CCDC188 in humans leads to the HTCA broken and separation of SUN5 and PMFBP1 into corresponding two parts.