Figure 4.

MPPED2 negatively modulates cell proliferation and migration of thyroid carcinoma cell lines. (A) qRT-PCR analysis performed on TPC-1 and FRO cell lines stably carrying MPPED2 or the corresponding empty vector (EV). Values were reported as relative expression ± SEM and were compared to EV, set equal to 1. t-test; * p < 0.05 (left panel). Immunoblot analysis confirming the expression of MPPED2. GAPDH and β-Actin were used to normalize the amount of loaded protein (right panel). (B) Cell growth analysis of TPC-1 and FRO stably carrying MPPED2 or EV. Cell number was evaluated at 24 h, 48 h, 72 h and 96 h after seeding. Values were obtained from three independent experiments performed in duplicate and data were reported as mean ± SEM. 2-way Anova-test followed by Bonferroni post-test; * p < 0.05; *** p < 0.001. (C) Representative colony assay performed on TPC-1 and FRO cell lines stably carrying MPPED2 or EV. (D) Representative cell cycle analysis of TPC-1-MPPED2 and TPC-1-EV cells. Cell number was reported on the y-axis while the percentage of propidium iodide (PI) incorporated was reported on the x-axis (left panel). Values shown in the right panel were obtained from three independent experiments. t-test; * p < 0.05 compared to EV. (E) Representative acquisition of migration assays performed on MPPED2 or EV transfected TPC-1 and FRO cells (magnification 40×) (left panel). Values obtained from three (TPC-1) or four (FRO) independent experiments were reported as mean ± SEM and compared to the EV, set equal to 1 (right panel). t-test; * p < 0.05. (F) qRT-PCR analysis to evaluate the expression of RP5-1024C24.1 after MPPED2 transfection. Data obtained from three (TPC-1) or five (FRO) independent experiments were reported as relative expression ± SEM and were compared to the EV, set equal to 1. t-test; p = ns.