QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 5. Effects of Nek2 on ectopic NIP2 proteins. (A) 293T cells were co-transfected with GFP-NIP2 and kinase-defective Nek2KHA5 (KD) or varying amounts of wild-type Nek2RHA1 (WT). Twenty-four hours later, cell lysates were prepared using a lysis buffer containing 1% NP40 and separated into the insoluble pellet (P) and soluble supernatant (S) by centrifugation. Equal amounts of pellet and supernatant proteins were subjected to immunoblot analysis using antibodies against GFP and the HA-tag for detection of ectopic NIP2 and Nek2 proteins, respectively. (B) Myc-NIP2 was co-transfected into HeLa cells with either Nek2RHA1 or Nek2KHA5. Co-immunostaining was carried out using NIP2 (green) and HA (red) antibodies. (C) HeLa cells that had been co-transfected with GFP-NIP2 and Nek2RHA1 were co-immunostained with antibodies against GFP (green) and Nek2 (red) or β-tubulin (red). DNA was stained with DAPI. Bars, 10 µm. (D) HeLa cells were transfected with Myc-NIP2^445-903. Twenty-four hours after transfection, nocodazole (100 µg/ml) was added for 2 hours and cells were immunostained with antibodies against NIP2 (green) and acetylated -tubulin (Ac-tubulin, red). (E) HeLa cells with GFP or GFP-NIP2^445-903 proteins were lysed in the presence of nocodazole or taxol and then fractionated into supernatant (S) and pellet (P). β-tubulin was used as a marker for fractionation.

Return to article