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. 2014 Sep 30;5:475. doi: 10.3389/fimmu.2014.00475

Figure 8.

Figure 8

Anti-apoptotic and anti-oxidant effects of EPO in SH-SY5Y cells may be regulated by miR-451 and miR-885-5p. Transfected and non-transfected cells were treated by staurosporine (STS) (25 nM) and 1 U/ml EPO. (A) DNA fragmentation was evaluated by cell death ELISA assay. (B) Apoptotic phosphatidylserine (PS) positive cells were stained by Annexin-V-FITC dye and visualized using immunofluorescence microscopy. Cells were also treated with CoCl2 (250 μM) and 1 U/ml EPO (C) and DNA fragmentation was analyzed by cell death ELISA assay. Transfection of miR-451 and miR-885-5p mimics reduced the anti-apoptotic effect of EPO in both STS and CoCl2 induced apoptosis. (D) Intracellular ROS production was quantified by using the CM-H2DCFDA method. Over-expression of miR-451 and miR-885-5p reversed anti-oxidant effect of EPO induced by CoCl2. STS; staurosporin, EPO; erythropoietin, CM-H2DCFDA; 2′,7′-dichlorodihydrofluorescein, acetyl ester. The data are presented as mean ± SE, n = 5 [*p < 0.05 compared to control and #p < 0.05 compared to toxic agents (STS or CoCl2) treated cells].