Fig 2: Quantification of PCNA, Pax6 and Tbr2 positive cells in the hippocampal dentate gyrus of wildtype and Thy1-aSyn mice. Double-immunofluorescence for PCNA (green) and Pax6 (red) and DAPI staining (blue) and merged images of 6-month-old wildtype [WT, (A)] and Thy1-aSyn [asyn, (B)] mice. Number of PCNA+ cells (C), Pax6+/PCNA+ cells (D) in the granule cell layer (GCL) and subgranular zone (SGZ) of 6- and 16-month (mo)-old wildtype (WT) and Thy1-aSyn (asyn) mice. Double-immunofluorescence for PCNA (green) and Tbr2 (red) and DAPI staining (blue) and merged image of 6-month-old wildtype [WT, (E)] and Thy1-aSyn [asyn, (F)] mice. Number of Tbr2 + /PCNA + cells (G) in the GCL and SGZ of 6 and 16 month-old (WT) and Thy1-aSyn (asyn) mice. Data represent mean ± SEM. (WT; 6 month-old, n = 6, 16 month-old n = 6; asyn, 6 month-old, n = 6, 16 month-old n = 5; Student’s t-test; *p < 0.05, ***p < 0.005). GCL, granule cell layer; ML, molecular layer; SGZ, subgranular zone; PL, polymorphic layer. Scale bars, 50 µm.

Fig 3: The association of Pol1, Pol2, Pol3, PCNA, Dpb11, Dpb2 and Rap1 with chromosomes during telomere dysfunction. All strains were cdc13–1, grown at 21°C, followed by 480 min at 27°C to induce telomere dysfunction. (A) Dynamics of ssDNA accumulation in sub-telomeres. (B–H) Dynamics of protein association with sub-telomeres. Proteins are indicated above each graph. Error bars are the standard deviation between three measurements. (I) Dynamics of ssDNA accumulation at the YER188W single gene locus. (J–P) As in (B–H), except that the protein association with YER188W was analyzed. (Q) ssDNA at the centromere-proximal locus PAC2. (R–X) Protein association with PAC2. Pol1 and Pol3 are tagged with HA. Dpb2 is tagged with Myc. Pol2, PCNA, Dpb11 and Rap1 are not tagged. (Y) The cell cycle distribution of cells analyzed in A–X.

Fig 4: Development of the germinal zones in the tree shrew neocortex. (A–D) Immunofluorescence for PCNA (yellow) and DAPI staining (blue) on 30 µm-cryosections of E32–P1 tree shrew neocortex. The top margin of the image corresponds to the transition zone SVZ/intermediate zone. Scale bars, 50 µm. VZ, ventricular zone; SVZ, subventricular zone; iSVZ, inner SVZ; oSVZ, outer SVZ. (E) Quantification of the VZ thickness of the E32–P1 tree shrew neocortex. (F) Quantification of the SVZ thickness of the E32–P1 tree shrew neocortex. (G) Quantification of the VZ and SVZ thickness of the E32–P1 tree shrew neocortex, expressed as percentage of the sum of VZ and SVZ. (H) Quantification of the iSVZ and oSVZ thickness of the E37 tree shrew neocortex, expressed as percentage of the sum of iSVZ and oSVZ. (E–H) Data represent mean ± SD and were obtained from two consecutive sections of two brains each.

Fig 5: Quantification of Tbr1 and NeuN positive cells in the hippocampal dentate gyrus of wildtype and Thy1-aSyn mice. (A,B) Immunofluorescence for Tbr1 (red) and DAPI staining (blue) and merged images of 6-month-old wildtype [WT, (A)] and Thy1-aSyn [asyn, (B)] mice. Number of Tbr1 + cells (C) in the granular cell layer (GCL) and subgranular zone (SGZ) of 6-month (mo)-old wildtype (WT) and Thy1-aSyn (asyn) mice. (D,E) Immunofluorescence for NeuN (red) and DAPI staining (blue) and merged images of 16-month-old wildtype [WT, (D)] and Thy1-aSyn [asyn, (E)] mice. Number of NeuN + cells (F) in GCL and SGZ of 6 month-old wildtype (WT) and Thy1-aSyn (asyn) mice. (G) Change (percent) in number of PCNA+, Pax6+/PCNA+, Tbr2+/PCNA+ cells and neurons between 6 and 16 month-old Thy1-aSyn (asyn) and wildtype (WT) mice. Data represent mean ± SEM. (WT; 6 month-old, n = 6, 16 month-old n = 6; asyn, 6 month-old, n = 6, 16 month-old n = 5; Student’s t-test). Scale bars, 50 µm.