Supplementary Figure 5: Suppression of C/D Box SnoRNAs Delays Leukemogenesis in vivo.
From: AML1-ETO requires enhanced C/D box snoRNA/RNP formation to induce self-renewal and leukaemia
(a) Sanger sequencing of Kasumi-1 CRISPR snoRNA knockout cells. Arrow shows the mutations induced by CRISPR/CAS9 in snoRNA loci. (b) to (c) SnoRNA knockout did not affect host gene expression. SnoRNA host gene RPL13A was analyzed both on mRNA level by RT-PCR (b) and on protein level by Western blot (c) in control and SNORD34 or 35A knockouts. Two single clones for each knockout were checked. (d) SnoRNA knockout did not affect total RNA content. Total RNA from 1 × 105 indicated Kasumi-1cells were eluted in 25 μl RNase-free H2O. Error Bar, mean ± s.d. of n = 5 measurement of 2 independent replicates. (e) Colony numbers formed by MV4-11 CRISPR snoRNA knockout cells, compared to control. (f) Kaplan–Meier survival analysis of NSG mice injected with snoRNA knockout or control MV4-11 cells. N = 12 (Control_gRNA), 7 (SNORD14D_gRNA) and 6 mice (SNORD35_gRNA), log-rank test. (g) Kaplan–Meier survival curve of NSG mice receiving 1 × 104 control (Ctr_gRNA, n = 12 mice) or SNORD43 CRISPR knockout (SNORD43_gRNA, n = 6 mice) MV4-11 cells. Error Bar, means ± s.d. of n = 3 independent experiments unless otherwise indicated. Unprocessed original scans of blots are shown in Supplementary Figure 7.
