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

. 2019 Mar 4;218(3):871–894. doi: 10.1083/jcb.201804183

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

© 2019 Dwivedi et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Figure 9. — Hook2 is required for p150^glued and MKLP1 interaction during cytokinesis. (A) Lysates of HEK293T cells synchronized in either G2 phase or cytokinesis were IP with control IgG or anti-MKLP1 antibody and IB with indicated antibodies. (B) Ratio of normalized band intensity (cytokinesis lane) of IP p150^glued and DIC to MKLP1 in A (n = 2). (C) Whole-cell lysates of HEK293T cells harvested during the cytokinesis stage were incubated with either MBP or MBP-tagged Hook2 N427 (WT, Q143A, and I150A) and IB for MKLP1. (D) Ratio of band intensity of pulldown to input Hook2 (WT/mutants) signal in C (n = 2). (E) Lysates of HEK293T cells treated with control or p150^glued siRNA and harvested during cytokinesis stage were IP with control IgG or anti-Hook2 antibody and IB with the indicated antibodies. (F) Ratio of normalized band intensity (control siRNA) of coIP proteins (as indicated) to MKLP1 in E (n = 2). (G) Lysates of HEK293T cells transfected with indicated siRNA and harvested during cytokinesis stage were tested for dynactin association with MKLP1. (H) Ratio of normalized band intensity (control siRNA) of IP p150^glued and DIC to MKLP1 in G (n = 3). (I) Representative images of HeLa cells ectopically expressing HA-tagged zebrafish Hook2 (HA-zHook2) costained for centrosomes, dynein, and dynactin. Bars, 10 µm; insets, 2 µm. (J) Lysates from asynchronous HEK293T cells treated with indicated siRNA and transfected with indicated plasmids were IP with control IgG or anti-DIC antibody and IB for p150^glued. (K) Ratio of normalized band intensity (control siRNA) of IP p150^glued to DIC in J (n = 2). (L) Representative differential interference contrast images of zebrafish embryos injected with either nontargeting control morpholino or indicated concentration of zHook2 morpholino immediately after fertilization and imaged at the indicated time. Each concentration of morpholino was injected in 100 fertilized embryos and monitored over time. Bars, 500 µm. Data represent mean ± SD (ns, not significant; ***, P < 0.001; Student’s t test). (M) Proposed model depicting the role of Hook2 in mediating formation of dynein–dynactin complex and targeting of MKLP1 to the spindle midzone.