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. 2017 Sep 5;114(38):10268–10273. doi: 10.1073/pnas.1704143114

Fig. 2.

Fig. 2.

Effects of DYRK1A overexpression or knockdown on NSC proliferation (A–D) and cyclin D1 expression (E–J). (A) Representative images and (B) quantitative analyses of NSCs expressing GFP or DYRK1A are shown. BrdU incorporation is visualized in red. (Scale bar, 10 μm.) (C) NSCs were transfected with lentiviral vectors expressing scramble shRNA (scr) and EGFP, DYRK1A #1 shRNA (hp#1) and EGFP, or Dyrk1A #2 shRNA (hp#2) and EGFP. Effective DYRK1A knockdown was observed 3 d posttransfection. (D, Left) Representative images of NSCs transfected with the indicated lentiviral vectors and labeled for BrdU incorporation (magenta). (Scale bar, 20 μm.) (Right graph) Ratios of BrdU double-positive cells to GFP-positive cells (% BrdU+&GFP+/GFP+) in DYRK1A-knockdown NSCs [F(4,12) = 4.06, P = 0.000149]. (E) Cyclin D1 expression in DYRK1A-expressing and DYRK1A kinase dead mutant-expressing cells. (F) DYRK1A coexpressed with Myc-cyclin D1 WT, T286A, T288A, or T286AT288A (2TA) mutants. The values at the bottom of the panels indicate protein levels normalized to control values. (E–H) Effects of NSC growth-promoting compounds #679, #688 (ALGERNON), and #690 on cyclin D1 expression. HEK293 cells (G) and NSCs (H) were treated with NSC growth-promoting compounds (#679, #688, and #690), a DYRK1A inhibitor (harmine), and structurally related negative control compounds (TG001 and TG009) (5 μM each) for the indicated durations or 24 h, respectively. *P < 0.05.