Identification of Cell Type-Specific Differences in Erythropoietin Receptor Signaling in Primary Erythroid and Lung Cancer Cells
Fig 7
Experimental validation of the cell type-specific CISH and SOCS3 mRNA parameters.
The parsimonious model was employed to predict the dynamics of CISH mRNA and SOCS3 mRNA upon treatment with a transcriptional inhibitor. The mRNA dynamics in CFU-E stimulated with 5 U/ml EPO alfa alone (black) or with transcriptional inhibition after 60 min (blue) was predicted. Additionally, the mRNA dynamics in H838-HA-hEPOR stimulated with 10 U/ml EPO beta alone (black) or with transcriptional inhibition after 30 min (blue) was predicted. Shadings surrounded by dotted lines depict uncertainty of the prediction. CFU-E cells were stimulated with 5 U/ml EPO alfa and either additionally treated with 1 μg/ml actinomycin D, to inhibit transcription, at 60 min (blue arrows) or left untreated. The H838-HA-hEPOR cells were either stimulated with 10 U/ml EPO beta alone (black) or additionally with 1 μg/ml actinomycin D at 30 min (blue arrows). The mRNA was extracted at the indicated time and the SOCS3 and CISH mRNA levels were measured with qRT-PCR. Experimental data are depicted as closed circles. The experiment was performed in triplicates and one representative example is shown.