Figure 4.
Knockout of SNORD42A reduces the 2′-O-methylation level of U116. (A) The 2′-O-methylation at different rRNA sites in 2 Kasumi-1 SNORD42A knockout (KO; vertical axis) and 3 control (horizontal axis) single clones is indicated. (B) Diagram for analysis of 2′-O-methylation with RT-PCR and different dNTP concentrations. (C) Mean Cq values for high and low dNTP concentrations in the 5 Kasumi-1 SNORD42A knockout and 6 control single clones. Data are shown as mean ± SD; n.s, not significant, ***P < .001, Student t test. (D) Relative 2′-O-methylation at 18S-U116 in 5 control and 6 SNORD42A knockout Kasumi-1 single clones. The data were normalized to the mean of the control. ***P < .001, Student t test. (E) Colony number formed by the same single clones that were used for 2′-O-methylation analysis in Figure 3D. Three hundred cells were seeded in methylcellulose. Data are shown as mean ± SD from 3 replicates, ***P < .001, Student t test. (F) Correlation between the number of colonies forming units and relative 2′-O-methylation in the single clones. Pearson correlation r = +0.80, P = .003. CFU, colony-forming unit.