Figure 3.

Treatment with cinnamon extracts up-regulates endogenous expression of ZFP36L1, down-regulates Stat5b levels, and inhibits erythroid differentiation of CD34+ HSCs. (A) Real-time PCR showing up-regulation of ZFP36L1 after treatment of Hel cells with 0.5 μg/μl cinnamon extracts for 24 h (SEM calculated on three experiments). (B) Real-time PCR performed on Hel cells treated with cinnamon extracts showing decreased expression of Stat5b and of the erythroid markers GATA1, LMO2, HbA1, and RhAG; CD44 monocyte differentiation marker expression is increased after treatment with cinnamon extracts; ACTB was used as control housekeeping gene. Results are provided in terms of Log₂ of relative quantity compared with the expression levels of the same genes in untreated Hel cells; error bars, SEM calculated on a set of three independent experiments (*p < 0.05; **p < 0.01). (C) Real-time PCR showing up-regulation of ZFP36L1 after treatment of CD34+ HSCs in liquid culture with 0.25 μg/μl cinnamon extracts for 24 h (SEM calculated on three experiments). (D) Western blot analysis of Stat5b levels in untreated CD34+ HSCs (lane 1) and in the same cells treated with 0.25 μg/μl for 48 h (lane 2). (E) Clonogenic assay performed on CD34+ HSCs in the absence (left) on in the presence (right) of cinnamon extracts. Cinnamon extracts were added directly to methylcellulose medium at a concentration of 0.25 μg/μl (SEM calculated on three independent experiments; *p < 0.05). (F) Real-time PCR showing down-regulation of Stat5b and erythroid markers in CD34+ HSCs treated with 0.25 μg/μl cinnamon extracts in liquid culture medium for 48 h; CD44 expression is up-regulated after treatment with cinnamon extracts; ACTB was used as control housekeeping gene. Data are provided in terms of Log₂ of relative quantity compared with the expression levels of the same genes in untreated CD34+ HSCs; error bars, SEM calculated on a set of three independent experiments (*p < 0.05; **p < 0.01).