Figure 3. MiR-125 regulates CD4 differentiation marker in T-ALL cells through targeting A20.

A. 1×10⁶ Jurkat-vector and Jurkat-A20 stable cell lines were incubated with CD4 and CD8 for 45min, washed, and then the cells were analyzed by flow cytometry. Jurkat-vector and Jurkat-A20 stable cell lines were collected. Cell lysates were prepared for Western blotting with an antibody against A20, and β-actin was used as a loading control. B. Jurkat cells were transfected with scrambled siRNA or A20 siRNAs. Forty-eight hrs after transfection, Cells were harvested and used for extracting RNA. qRT-PCR was performed to validate the expression of CD4 in Jurkat with depletion of A20 by siRNA. GAPDH was used as an internal control and for normalization of the data. Columns represent the mean of three independent experiments; bars represent SE. ***, p<0.001. C and D. 1×10⁶ Jurkat-vector and Jurkat-miR-125b, Jurkat-miR-125b transfected with A20, T2 vector, T2-miR-125b, T2-miR-125b transfected with A20 cell lines were incubated with CD4 and CD8 antibodies for 45min, washed, and then the cells were analyzed by flow cytometry. bars represent SE. **, p<0.01.