Fig. 5. DYRK1A inhibition regulates proliferation in a cell-dependent manner.

A Genomic profiles of glioblastoma stem cell lines HW1, RN1, MMK1 and RKI. B Immunoblot analysis (72 h) and nuclear staining quantification (7 days) of cells transfected with scramble (si Ctr) and DYRK1A-targeting (si DYR) siRNAs. Scale bar: 10 μm. C Manual counting of cells transfected with si Ctr and si DYR for 7 days. D Representative images and quantification of colonies in cells transfected with si Ctr and si DYR for 14 days. E RT-PCR analysis of cell cycle genes in cells transfected with si Ctr and si DYR for 7 days. F Flow cytometry quantification of cells in G1/G0, S and G₂/M phases following transfection with si Ctr and si DYR for 3 days. G Nuclear staining quantification of cells treated with DYRK1A inhibitor ALGERNON (ALG) for 7 days. H Representative images and quantification of colonies in cells treated with ALG for 14 days. I Immunofluorescence images and quantification of mitotic figures in cells treated with ALG for 7 days. Cells were stained using antibodies against Ki67 (pink) and tubulin (green). DAPI was used to visualize cell nuclei (blue). Scale bar: 5 μm. All bar graphs represent mean ± SEM from at least three independent experiments (two-tailed, unpaired t-test, *P < 0.05, **P < 0.01, ***P < 0.001).