Long-term treatment with miR-29b suppressed proliferation of leukemic cells in vivo. (A) RNA-seq from purified splenic CD19⁺ B lymphocytes from the mice treated with 2A2-miR-29b-ILP or 2A2-control scramble-ILP. Differential expression of genes revealed at least 233 differentially expressed genes between miR-29b vs scramble control (128 genes decreasing and 105 genes increasing). (B) A total of 128 of the 233 differentially expressed genes represent cellular growth and proliferation. (C-D) Cell cycle analysis of CD19⁺ B cells from the in vivo 2A2-miR-29b ILP–treated group compared with the control 2A2-scramble-ILP–treated group. Panel C shows representative data of vehicle, 2A2-scramble-ILP2 (SC), and 2A2-miR29b ILP (mR-29b) treated groups, and panel D presents the summarized data (P = .0002; n = 4). Panel D also shows increased apoptotic cells in the 2A2-miR29b-ILP treatment group (scramble, 2.55 ± 0.8%; miR-29b, 16.9 ± 3.1%). (E) Validation of decreased expression of SP1, ZAP70, and CTLA-4 mRNA and increased expression of CDKN1a/p21 mRNA in the post–2A2-miR-29b-ILP treatment group confirmed by differential expression of genes.