Figure 2.

Induction of intron retention / spliceosome modulation mediated by FD-895 and PLAD-B. (A) RT-PCR was used to access spliced (S) and unspliced (U) isoforms of DNAJB1 and GAPDH in CLL cells. (B) IR was evaluated by RT-PCR for DNAJB1, RIOK3 and GAPDH (used as loading control) in CLL cells after treatment. (C) Quantitative real-time-PCR analysis of CLL cells after treatment to evaluate levels of unspliced mRNA for DNAJB1. GAPDH was used for normalization. (D) Intron retention for DNAJB1 and (E) RIOK3 was evaluated by quantitative real-time-PCR in CLL and normal B cells using specific primers that allowed detection of intron-containing regions (Online Supplementary Table S1). The untreated controls (CLL cells or NBC) for DNAJB1 and RIOK3 were set to a value of 1. GAPDH was used as a control for normalization.