Supplementary Figure 6: SYNCRIP post-transcriptionally controls HOXA9 expression.

(a) MOLM13 human myeloid leukemia cells carrying the MLL-AF9 translocation overexpressing MSI2 were immunoprecipitated for endogenous MSI2 and SYNCRIP. (b) Immunoblots showing protein expression of HOXA9, MYC and IKZF2 upon SYNCRIP knockdown in dsRed MLL-AF9 cells 3 and 4 d after transduction. Actin serves as a loading control. (c) Immunoblots showing protein expression of HOXA9 and MYC upon SYNCRIP knockdown in MOLM13 cells after 2 d of puromycin selection (4 d after transduction). (d,e) qPCR measuring the mRNA expression of Syncrip, Hoxa9, Myc and Ikzf2 in dsRed tertiary MLL-AF9 leukemia cells 3 and 4 d after transduction. (f) Immunoblots showing protein expression of HOXA9, MYC and SYNCRIP upon MSI2 knockdown in dsRed MLL-AF9 cells at 4 d after transduction. (g) qPCR measuring mRNA expression of Syncrip, Hoxa9, Myc and Ikzf2 upon MSI2 knockdown in dsRed MLL-AF9 cells at 4 d after transduction. (h) qPCR measuring mRNA expression of Syncrip, Hoxa9, Myc and Ikzf2 in RN2 cells 24 h after induction. (i) qPCR measuring mRNA expression of SYNCRIP, HOXA9, MYC and IKZF2 in human MOLM13 leukemia cells 3 d after transduction. (j) qPCR measuring mRNA expression of SYNCRIP, HOXA9, MYC and IKZF2 in human MOLM13 leukemia cells 4 d after transduction. (k) Normalized total RNA levels in dsRed MLL-AF9, RN2 and MOLM13 cells upon SYNCRIP depletion. (l) mRNA stability of Syncrip, Hoxa9, Myc and Ikzf2 in dsRed cells transduced with control and SYNCRIP shRNAs 4 d after transduction. (m) mRNA stability of Syncrip, Hoxa9, Myc and Ikzf2 in RN2 cells expressing Cas9-EV or gRNA1 and gRNA3 targeting Syncrip after 24 h of Dox induction. Actin served as a loading control. Actb served as a control housekeeping gene. All data represent the means + s.e.m. of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed t test.