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Fig. 4.
Dyrk1a localizes to mitotic spindles and is required for cell cycle progression and survival. (A-B‴) Stage 10 embryos stained for Dyrk1a (green), α-Tubulin (microtubules, magenta) and DAPI (DNA, blue). White arrowheads indicate Dyrk1a puncta near mitotic spindle. (C,D,G,H,G′,H′,J,J′,K,K′) Dorsal view of X. tropicalis tadpoles treated with DMSO (top row) or 1.25 µM Dyrk1a inhibitor harmine (bottom row). (C,D) Antibody staining for PCNA (proliferating cell nuclear antigen, S-phase marker, green). Telencephalon region is shown in the inset. (E) Quantification of first ventricle area using PCNA staining. (F) Quantification of the ratio of neural progenitor cells (NPCs, PCNA area) to the area of differentiated neurons in each telencephalon (log scale). (G,H) Antibody staining for pHH3 (phospho-histone H3, M-phase marker, magenta). (G′,H′) High-magnification views of telencephalon regions from G and H. (I) Quantification of pHH3-positive cell number per telencephalon. See also Fig. S4. (J,K) Antibody staining for CCP3 (cleaved caspase 3, cell death marker, yellow). (J′,K′) High-magnification view of telencephalon from J and K. (L) Quantification of CCP3-positive cell number per telencephalon. Scale bars: 200 µm. Whiskers are maximum and minimum values, boxes show the interquartile range and the line is the median. Every point is the value for one animal. P-values were calculated using non-parametric Mann–Whitney rank sum tests. |
