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The cytoprotective effect of asialo-rhuEPOP and Western blot of JAK2 and caspase 3.

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posted on 2013-10-04, 02:51 authored by Farooqahmed S. Kittur, Mamudou Bah, Stephanie Archer-Hartmann, Chiu-Yueh Hung, Parastoo Azadi, Mayumi Ishihara, David C. Sane, Jiahua Xie

N2A cells were treated individually with PBS containing 0.1% BSA (vehicle control), 1 µM STS, 1 µM STS+20 U/ml asialo-rhuEPOP (rhuEPOP) or 1 µM STS+20 U/ml rhuEPOM. (A) Cytotoxicity was measured by LDH assay after 12 h treatment. Each experiment had six replicates. All data plotted are the average of three independent experiments ± SD. *, P<0.05. (B) Western blot of activated JAK2 and caspase 3 in cell lysates prepared from cells treated by STS and rhuEPO for 3 and 6 h. For detection of p-JAK2 and JAK2, the blot was probed with anti-p-JAK antibody first followed by stripping the blot and re-probing with anti-total JAK2 antibody. Active caspase 3 was detected using an anti-caspase 3 antibody, which also cross-reacts with procaspase 3. β-actin was used as internal control.

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