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. 2016 Mar 14;126(4):1495–1511. doi: 10.1172/JCI80046

Figure 9. Expression of IGF2BP3 RNA-binding domain mutants in vivo and in vitro.

Figure 9

(A) Schematic of IGF2BP3 with its binding domains and the respective mutants (KH and RRM). (B) Time course of normalized engraftment to MIG in PB between 4 and 20 weeks after transplant. (C) FACS of PB done at 4 weeks after BMT, showing CD45.2 and GFP positivity (one-way ANOVA followed by Bonferroni’s test; ***P < 0.001). (D) B cells in PB 16 weeks after transplant. (E) Myeloid cells in PB 12 weeks after transplant. n = 8 for all groups. Mutant BMT experiment was completed twice for validation. (F) Western blot confirmed expression of IGF2BP3 (64 kDa), KH (47 kDa), and RRM (22 kDa) proteins in 7OZ/3 using anti-T7 (top panel) and anti-IGF2BP3 (bottom) antibodies. Actin used as a loading control. (G) Luciferase assay shows increased luciferase activity for the MYC 3′UTR when cotransfected with hI3 and decreased luciferase activity for the MYC 3′UTR when cotransfected with KH and RRM mutants (t test hI3, K,H and RRM; ***P < 0.001; ****P < 0.0001). Experiment was completed 3×. Data represent mean ±SD. hI3, human IGF2BP3; PB, peripheral blood.