NOTCH2-FL and FLT3-FL and their novel splice variant expression frequencies in AML patients and their association with disease status. (A-B) These figures display overall expression patterns of (A) NOTCH2 and (B) FLT3 FL transcripts and their splice variants. On the figures, the x-axes display patient and normal donor samples, and the y-axes display relative fluorescent units (RFU). PCR product RFU = log₂RFU. RFU is a unit of measurement calculated relative to the size standards included in each reaction. For relative level determination, product levels were kept below 3500 RFU, and size standard levels were within 500 to 800 units as recommended by the manufacturer. All calculations and an instrument calibration were done according to the Life Technologies recommendations. We note that in some patients, FLT3-FL expression is lower than the cutoff and not displayed on the graph. (C-E) These figures display overall expression patterns of NOTCH2-FL and FLT3-FL transcripts and their splice variants over the course of the disease in 17 patients. Patients are numbered from 1 to 17; patients samples were taken (C) at diagnosis and relapse (patients 1-8), (D) at diagnosis and remission (patients 9-13), or (E) over the course of refractory disease (patients 14-17). A total of 35 samples were collected including 14 samples obtained at diagnosis: Diag 1 to 13 and Diag 17; 10 samples obtained at relapse: Rel 1 to 8, Rel-14-1, and Rel-14-2 (first and second relapses); 5 samples obtained at remission: Rem 9 to 13; 6 samples obtained during regular visits over the course of refractory disease: Pers 15 to 17 (samples taken at first and second visits are marked as −1 or −2). On the panels, NOTCH2-FL and FLT3-FL and their splice variant transcripts expression levels are presented as log₂(PCR product RFU) and shown in a green color-coded scale.