Effects of treating Jeko-1 cells with antisense oligonucleotides (ASOs) and scrambled oligonucleotides (SCOs). (A) Expression of MiR-17 and miR-20a of Jeko-1 cells after targeting by ASOs or SCOs (AS-17+20 and SC-17+20, respectively). Expression of miR-19a of Jeko-1 cells treated with antisense miR-19a (AS-19) and scrambled miR-19a (SC-19) is also shown in this figure. (B) RQ-PCR analysis of CDKN1A expression in Jeko-1 cells treated with SC/AS-19, SC/AS-20, SC/AS-17, or SC/AS-17+20. CDKN1A mRNA levels in each sample were normalized to the corresponding level of G6PDH mRNA. The significance of differences between SC and AS was evaluated using Student t test. Error bars represent the mean (± SD) of 3 independent experiments. (C) RQ-PCR analysis of TP53INP1 expression in Jeko-1 cells treated with SC-17+20 or AS-17+20a. Error bars represent the mean (± SD) of 3 independent experiments. (D) Northern blot analysis (NB) of CDKN1A expression and Western blot analysis (WB) of p21, CDK6, CDK2, p27, Cyclin D1 (CCND1), and tubulin expression in Jeko-1 cells treated with AS-17+20 or SC-17+20. (E) Cell-cycle analysis of Jeko-1 cells treated with SC-17+20 or AS-17+20. Average (n = 3) percentage of SC-treated cells in G1 phase: 56.9%, S phase: 37.6, and G2 phase: 5.5%; and of AS-treated cells in G1 phase: 71.3%, S phase: 22.7%, and G2 phase: 6.0%. X-axis represents DNA content; y-axis, number of cells. Horizontal bars represent cell-cycle phases.