Figure 4

miR-101 differentially regulates EZH2 at mRNA and protein levels in subset 1 versus 4 CLL cases. (A) EZH2 mRNA levels in subsets 1 and 4 (four and four cases, respectively). The y axis represents relative expression (2^−ΔCt). The box-and-whisker plots repesent median with minimum and maximum values. EZH2 is upregulated in subset 1 cases (fold difference in expression of 2.3; p ≤ 0.05). (B) Correlation and linear regression between miR-101 and EZH2 mRNA levels. ΔCt values were used and the correlation is statistically significant (p ≤ 0.05). The slope is negative, which means that when miR-101 expression is upregulated, EZH2 expression is downregulated (r = −0.75). (C) Protein levels of EZH2 in subsets 1 and 4 (six and eight cases, respectively). EZH2 protein levels are higher in subset 1 (p < 0.005). (D) Correlation and linear regression between miR-101 relative expression (2^−ΔCt) and EZH2 protein levels (r = −0.7, p < 0.05). (E) In subset 1 CLL cells, miR-101 mimic induces downregulation of EZH2 protein levels compared with untransfected cells and cells transfected with negative control. (F) In subset 4 CLL cells, miR-101 inhibitor induces EZH2 protein upregulation compared with untransfected cells or cells transfected with negative control.