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. 2015 Jul 23;34(19):2424–2440. doi: 10.15252/embj.201490831

Figure 5.

Figure 5

Phosphorylation of NDE1 by CDK5
  1. A HEK293T cells were transfected with indicated plasmids, and total cell lysates were separated through SDS–PAGE containing 50 μM Phos-tag™ reagent. Two asterisks indicate phosphorylated species in two sites, whereas one asterisk indicates phosphorylated species in one site.
  2. B Quantification of NDE1 phosphorylation in two sites (shown by two asterisks in A) co-transfected with indicated kinases and activators.
  3. C Mono-phosphorylation of endogenous NDE1 in RPE1-hTERT cells. Cell lysates from HEK293T cells transfected with Flag-tagged NDE1 (F-NDE1, lane 1) or F-NDE1 + CDK5 + p25-GFP (lane 3) or untransfected RPE1-hTERT cells (lane 2) were separated by SDS–PAGE containing 50 μM Phos-tag™ reagent and immunoblotted with rabbit α-NDE1.