MPLW515L-Expressing Cells Show Hyper-Responsiveness to TPO

<div><p>(A) 32D MPLWT cells and 32D MPLW515L cells were cultured in triplicate at an initial concentration of 1 × 10<sup>5</sup> cells/mL, in RPMI/10% FBS containing TPO 2 ng/mL, 1 ng/mL, 0.1 ng/mL, 0.01 ng/mL, 0.001 ng/mL, or in the absence of TPO for four days, and then cell numbers were assessed. Error bars denote the standard deviation for each sample measured in triplicate. </p> <p>(B) UT7 MPLWT cells and UT7 MPLW515L cells were cultured in triplicate, at an initial concentration of 1 × 10<sup>5</sup> cells/mL, in IMDM/10% FBS containing TPO 5 ng/mL, 1 ng/mL, 0.1 ng/mL, 0.01 ng/mL, 0.001ng/mL, or in the absence of TPO for six days, and then cell numbers were assessed. Error bars denote the standard deviation for each sample measured in triplicate. </p> <p>(C) 32D MPLWT or 32D MPLW515L cells were deprived of cytokine overnight and then stimulated with TPO 5 ng/mL, 1 ng/mL, 0.1 ng/mL, 0.01 ng/mL, or without TPO for seven minutes and then analyzed by Western blot for phosphorylation of JAK2 and STAT3, demonstrating increased JAK2/STAT3 phosphorylation in response to TPO in 32D MPLW515L cells as compared with 32D-MPLWT cells.</p> <p>(D) Phosphorylation of JAK2, TYK2, STAT3, STAT5, AKT, and ERK in response to seven-minute stimulation with TPO (50ng/mL). 32D MPLWT or 32 MPLW515L cells were deprived of cytokine overnight and then stimulated with TPO 50 ng/mL for seven minutes and then analyzed by Western blot for phosphorylation of JAK2, TYK2, STAT3, STAT5, AKT, and ERK, demonstrating increased phosphorylation of these proteins in response to TPO in 32D MPLW515L cells as compared with 32D MPLWT cells.</p></div>

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