Functional Interactions between BM88/Cend1, Ran-Binding Protein M and Dyrk1B Kinase Affect Cyclin D1 Levels and Cell Cycle Progression/Exit in Mouse Neuroblastoma Cells
Figure 8
Interaction of RanBPM with Dyrk1B or Cend1 affects negatively Neuro 2a cell differentiation.
(a) Neuro 2a cells were transiently single- and double-transfected with: Dyrk1B, pCAG-Cend1-IRES-GFP, RanBPM, RanBPM and Dyrk1B, RanBPM and pCAG-Cend1-IRES-GFP expression plasmids, as indicated. Cells were allowed 16 h for protein expression and were then induced to differentiate by treatment with 20 μM RA/ 2% FCS for 48 h after which they were fixed and immunofluorescently labeled for Dyrk1B, Cend1, RanBPM and the neuronal differentiation marker βIII-tubulin (Tuj1 antibody). Note that Cend1 is co-expressed with GFP (v, vi) which serves for visualization of Cend1-positive cells (vi, xviii). Scale bar: 40 μm. (b) Quantification of the mean neurite length in single and double-transfected cells (as indicated) as well as in non-transfected cells in each group. Different groups were compared by one-way ANOVA followed by Student's t-test. ***Student’s t-test: p<0.001, **: p<0.01, *: p<0.05, n= 3. Error bars represent SEM. #: represents statistically significant differences between transfected and non-transfected cells in the same group, while *: represents statistically significant differences among different groups.