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

Figure 2.

Figure 2

Upregulation of NDE1 in cells lacking FBW7
  1. A Expression levels of FBW7, NDE1, Aurora A kinase, and CDK5 in wild-type and FBW7-depleted RPE1-hTERT cells in exponentially growing un-trans-fected cells (untreated, lane 1) or 24-h serum-starved cells transfected with a scrambled siRNA (control siRNA, lane 2) or separately with two FBW7-specific siRNAs (siRNA#1 or siRNA#2) (lanes 3 and 4).
  2. B Quantification of fluorescence intensity ratio of NDE1/γ-tubulin (green/red) signals at the centrosome in 0-, 12-, and 24-h serum-starved transfected cells with a scrambled (control) or FBW7-specific siRNA (FBW7 siRNA#1). Fluorescence intensity of coinciding green or red pixels within the box area (inset) was measured. The range of fluorescence intensity per pixel in a box was 0–255 (total number of cells from three independent transfections is indicated on graph). Data represent mean ± SEM. One-way ANOVA followed by Newman–Keuls post-test was used to determine significant difference among groups. ***P < 0.005, ns indicates no significance.
  3. C Upregulation of steady-state levels of NDE1 in DLD-1 FBW7−/− cells. Lysates prepared from exponentially growing (lanes 1 and 5) and serum-starved FBW7−/− (lanes 2–4) or wild-type DLD-1 cells (lanes 6–8) were immunoblotted with rabbit α-NDE1 (upper panel), mouse α-C-MYC (middle panel), or α-β-actin (lower panel).