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Analysis of EphA2 protein and RNA expression on GFP-control, MLL-AF9 and BCR-ABL bone marrow and spleen.

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posted on 2015-06-17, 03:16 authored by Sara Charmsaz, Kirrilee Beckett, Fiona M. Smith, Claudia Bruedigam, Andrew S. Moore, Fares Al-Ejeh, Steven W. Lane, Andrew W. Boyd

(A) EphA2 mRNA expression levels relative to 18sRNA in bone marrow of GFP-control, MLL-AF9 and BCR-ABL mice, analyzed per 1000000 18s RNA using RT-PCR. Significantly higher EphA2 expression (P = 0.0267) in the MLL-AF9 bone marrow compared to GFP-control mice. (B) Representative flow cytometric overlay analysis of the EphA2 expression in GFP-control, MLL-AF9 and BCR-ABL bone marrow. (C) Flow cytometric analysis of the EphA2 expression in bone marrow of GFP-control, MLL-AF9 and BCR-ABL mice measured as mean fluorescent intensity, showed significantly higher EphA2 expression on MLL-AF9 bone marrow compared to GFP-control bone marrow (P <0.0001) and compared to BCR-ABL leukemic bone marrow (P = 0.0019) (n = 5 GFP-control, n = 4 MLL-AF9, n = 3 BCR-ABL, 1 biological replicates, 2 technical replicates). (D) Flow cytometric analysis of the EphA2 expression in spleen of GFP-control, MLL-AF9 and BCR-ABL mice measured as mean fluorescent intensity, showed significantly higher EphA2 expression in spleen of the MLL-AF9 mice compared to GFP-control mice (P = 0.0143) and higher expression compared to BCR-ABL leukemic mice (n = 5 GFP-control, n = 4 MLL-AF9, n = 3 BCR-ABL, 1 biological replicates, 2 technical replicates). Each dot corresponds to one individual mouse. The data represent the mean ± SEM. Unpaired t test was performed for statistical analyses.