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Fig. 5 | Molecular Cancer

Fig. 5

From: Long noncoding RNA ZFAS1 promoting small nucleolar RNA-mediated 2′-O-methylation via NOP58 recruitment in colorectal cancer

Fig. 5

ZFAS1 promotes rRNA G3878 and G4593 2′-O-methylation by targeting NOP58 protein. a, CLIP database showing the motif of ZFAS1 targeting NOP58 predicted by online software (http://lulab.life.tsinghua.edu.cn/postar/index.php). b, Bioinformatics online software predicting the specific binding sequence and sites of ZFAS1 secondary structure and NOP58 protein (http://www.tartaglialab.com/). c, MOE multi-functional docking platform showing the specific docking sites between ZFAS1 tertiary structure and NOP58 protein. d, RNA pull-down followed by western blot showed in vitro binding of the ZFAS1-Wild, ZFAS1-Mutant, and antisense RNA probes with NOP58 protein after ZFAS1 silencing in SW620 cells. The biotin labeled probes are presented in Table S6. e and f, The NOP58 expression after overexpressing or knockdown NOP58 by qRT-PCR assay (e), and ZFAS1 rescued NOP58 mRNA expression after co-transfected ZFAS1 and NOP58 (f). g, Western blot assays detected the NOP58 protein expression after overexpressing, knocking down NOP58, as well as co-transfected ZFAS1 and NOP58 vectors. h and i, The 28S rRNA G3878 and G4593 2′-O-Me activities after overexpressing or knockdown NOP58 in SW620 and HCT116 cells by RTL-P (h), and ZFAS1 rescued the 2′-O-Me activities after co-transfected ZFAS1 and NOP58 (i).*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

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