Validation of interferon-stimulated genes in immortalized megakaryocyte cell line (imMKCL). (A) Immunoblot analysis of total SRC expression on day 0, 2 and 4 megakaryocytes (MK) (left). GAPDH was used as loading control. Quantification of SRC expression in triplicate immunoblots and statistical analysis using one-way ANOVA with multiple comparisons (right). (B) Quantitative real-time polymerase chain reaction (qRT-PCR) validation showing relative expression of SRC, IFIT1, MX1, OAS2, IFIT3 and ISG15 on day 0, 2 and 4. Statistical analysis was performed using one-way ANOVA with multiple comparisons. (C) qRT-PCR validation showing relative expression of SRC, IFIT1, MX1, OAS2, IFIT3 and ISG15 on day 2 MK transfected with empty vector, WT-SRC or E527K-SRC in pSecTag- Hygro expression vector (triplicated transfection experiment). Statistical analysis was performed using one-way ANOVA with multiple comparisons. (D) qRT-PCR validation showing relative expression of SRC, RUNX1, IFIT1, MX1, OAS2, IFIT3 and ISG15 in MK from a control and the patient carrying the E527K SRC variant (3 technical repeats). Statistical analysis was performed using one-way ANOVA with multiple comparisons.