Figure 1

In-vitro experiments. (a) miR-17 expression level in primary UM IGHV CLL cells left unstimulated (control) or stimulated with CpG-ODN (CpG) and in MEC-1 cell lines, as investigated by quantitative real-time polymerase chain reaction (qRT-PCR). Data represent mean ± SEM. (b) Expression of miR-17 in MEC-1 cells transfected with antagomiR17 or scrambled control. miR-17 expression was evaluated by qRT-PCR at different time-points (2 and 4 days). Data represent mean ± SEM of three replicates. P values (Student’s t-test) for each time-point are shown. *P < 0.05 (antagomiR17 versus scrambled control). (c) Expression of TRIM8, ZBTB4, and TP53INP1 in MEC-1 cells transfected with antagomiR17 or scrambled control. Gene expression was evaluated by qRT-PCR at different time-points (2 and 4 days). Data represent mean ± SEM of three replicates. P values (Student’s t-test) for each time-point are shown. *P < 0.05 (antagomiR17 versus scrambled control). (d) Effects of antagomiR17 transfection on TP53INP1, TRIM8 and ZBTB4 protein levels in MEC-1 cells. Protein expression levels were measured by Western blot experiment. Lower panel. Relative TP53INP1, TRIM8 and ZBTB4 protein expression levels of MEC-1 cells transfected with antagomiR17 or scrambled control assessed by Western blot. β- Actin levels were used as loading control in all cases. Upper panel. In all graphs values are represented as mean fold expression with respect to transfection with scrambled control. Data represent mean ± SEM of four replicates. P values (Student’s t-test) for each time-point are shown. *P < 0.05 (antagomiR17 versus scrambled control). (e) Proliferation of MEC-1 cells transfected antagomiR17. MEC-1 cells were transfected with antagomiR17 or scrambled control and counted once a day for four days. Dotted line indicates scrambled control transfected cells and solid line indicates antagomiR17 transfected cells. P value (Student’s t-test) is shown. Data represent mean ± SEM of three biological replicates.