Figure 6

CDK6 impairs monocyte/macrophage differentiation of CD34⁺ HSPCs.

(a) The expression of CDK6 was detected by western blot during PMA-induced THP-1 and HL-60 cells as well as monocytic differentiation of CD34⁺ HSPCs. (b) Western blot analysis of CDK6 expression in monocyte/macrophage induction cultures of CD34⁺ HSPCs infected with lenti-CDK6 and lenti-ctrl. (c) Expression of the monocytic differentiation marker CD14 was evaluated by flow cytometry after the infection and induction of CD34⁺ HSPCs. A representative experiment is presented in the left and the results from two independent experiments were statistically analyzed and presented as mean ± SD in the right. The red line and the black line indicate untreated cells and CD14 antibody stained cells respectively. (d) Real-time PCR analysis of CD14 mRNA expression at 17 days of differentiation of CD34⁺ HSPCs infected with lenti-CDK6 and lenti-ctrl. (e) May-Grünwald Giemsa staining of the infected and differentiation-induced CD34⁺ HSPCs. The cells were observed under ×40 magnification. The proportion of differentiated monocyte/macrophages was counted. Data are represented as mean ± SD. *P < 0.05 and **P < 0.01, Student’s t-test.