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. 2017 Oct 5;102(12):2015–2020. doi: 10.3324/haematol.2016.158857

Figure 2.

Figure 2.

Effect of lenalidomide on erythropoietin elaboration in the HepG2 cell line. (A) HepG2 cells were treated with 1 μM lenalidomide for 30 min prior to addition of S100A9 (1 μg/mL), TNFα (10 ng/mL) or IL-1β (10 ng/mL). Supernatant erythropoietin (EPO) concentration was determined by ELISA (B) HepG2 cells were treated with 0.1, 1 or 10 μM lenalidomide. Supernatant TNFα concentration was determined by ELISA (C) HepG2 cells were treated with 1 μM lenalidomide 30 min prior to addition of S100A9 (1 μg/mL) or TNFα (10 ng/mL). Quantitative polymerase chain reaction for IL10 mRNA expression was performed 24 h after treatment. GAPDH was used as an endogenous control and results are expressed as IL10 mRNA relative expression. (D) HepG2 cells were treated with 1 μM lenalidomide 30 min before addition of S100A9 (1 μg/mL) or TNFα (10 ng/mL). NF-κB protein was visualized by western blot 24 h after treatment. Rho GDI and lamin A/C were used as the loading controls for cytoplasmic and nuclear fractions, respectively. *mean P≤0.05.