Figure 3.
D2-type dopamine receptors regulate the transcription profile and cell cycle status of LSK cells. Unsupervised clustering (A) and principal components analysis (B) of RNA-seq results from Drd-DKO and WT LSK cells. (C) Bland-Altman plot showing 847 upregulated and 965 downregulated genes in Drd-DKO LSK cells relative to control at P <0.05 significance level. Red dots indicate differentially expressed genes. (D) Gene ontology analysis indicating the top downregulated pathways in Drd-DKO LSK cells. The yellow highlighting denotes cell cycle and mitosis-related pathways. (E) KEGG analysis indicating downregulation of the cell cycle pathway in Drd-DKO LSK cells. (F) Percentage of LSK cells with diploid (2n) or tetraploid (4n) DNA content after treatment with vehicle (n = 6) or haloperidol (n = 6). The P value was calculated using the Student t test. (G) Maximum intensity projections showing EdU incorporation in sections of distal long bone and, at higher magnification, in metaphysis after haloperidol treatment (left panels). Quantification of the number of EdU+ cells in metaphysis (area = 1 mm2) (right panel). Vehicle = 4; haloperidol = 4. (H) FACS-based quantification of EdU+ LSK cells and p-Histone H3+ EdU+ LSK cells after haloperidol treatment. The P value was calculated using the Student t test. Vehicle = 7; haloperidol = 7. Heat map of selected cell cycle (I), metaphase-anaphase transition (J), spindle/centrosome function (K), and DNA mismatch repair–related genes (L) in Drd-DKO and WT LSK cells.