Gene expression and RQ-PCR analyzes. (A) Western blot analysis of Bim and Bcl2 from Raji and SUDHL4 cells transfected with miR-17-19b-1 (Raji-m1 and Raji-m2/SUDHL4-m1 and SUDHL4-m2) or empty vector (Raji-c1 and Raji-c2/SUDHL4-c1 and SUDHL4-c2). Fold changes are shown below the gel normalized to tubulin, which was assigned a value of 1.00. Bim expression in Raji-m1 cells is 0.56 times lower than in Raji-c1 cells. Bcl2 expression in Raji-m1 is 1.43 times higher than in Raji-c1. (B) RQ-PCR analysis of CDKN1A and TP53INP1 expression in SUDHL4 and Raji cells transfected with miR-17-19b-1 (■) or empty vector (□). G6PDH served as an internal control, and the level of each gene's mRNA in each of the samples was normalized to the corresponding G6PDH mRNA level. The significance of the difference between SUDHL4 cells with (SUDHL4-m1) and without (SUDHL4-c1) miR-17-19b-1 transfection is shown above the bars. Values of P were calculated using Student t test. Error bars represent the mean (± SD) of 3 independent experiments. (C) Western blot analysis of p53, p21, CDK4, and CDK6 expression in Raji and SUDHL4 cells transfected with miR-17-19b-1 (Raji-m1 and S4-m1, respectively) or empty vector (Raji-c1 and SUDHL4-c1, respectively). Fold changes are shown below the gels. (D) Luciferase reporter assays of CDKN1A expression in Rat-1 cells transfected with miR-17-19b-1. MiR-17 expression in Rat-1 cells transfected with miR-17-19b-1 or empty vector (control) is shown on the right. The significance of differences between pGL3-CDKN1A 3′UTR wild-type and pGL3-CDKN1A was evaluated using Student t test. Error bars represent the mean (± SD) of 3 independent experiments.