Figure 2.
mRNA expression of POT1, TIN2, RAP1 and TPP1 in granulocytes derived from patients with myeloproliferative neoplasm (MPN) and JAK2V617F-dependent POT1 upregulation. (A and D) mRNA levels of each shelterin factor were analyzed with quantitative PCR and expressed arbitrarily based on the ratio of the target and β2-microglobin CT values. (A) POT1. (B) TIN2. (C) RAP1. (D) TPP1. (Left) Comparison between all MPNs and healthy controls, and right panels: Comparison between each subtype of MPNs and healthy controls. (E and F) Protein expression analyzed with western blot and signal quantified in ImageJ, (E) POT1, (F) TPP1. (G) Original protein blots of POT1, TIN2 and TPP1, and β–actin as loading reference. (H) POT1 mRNA in granulocytes from MPN patients with and without JAK2V617F mutation. (I) The correlation between JAK2V617F allele burdens and POT1 mRNA expression. (J–K) POT1 mRNA expression and protein expression in HEL erythroleukemia cell line (JAK2V617F+/+) cells treated with the JAK2-inhibitor LY2784544. Cells were exposed to LY2784544 (3 μM) for 24 hrs, RNA and protein was extracted and POT1 levels were then determined using qPCR and Western blotting, respectively. HC: healthy controls; MPN: myeloproliferative neoplasm; ET: essential thrombocythemia; PV: polycythemia vera; and PMF: primary myelofibrosis.