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Fig. 2 | Molecular Cancer

Fig. 2

From: Epigenetic inactivation of the 5-methylcytosine RNA methyltransferase NSUN7 is associated with clinical outcome and therapeutic vulnerability in liver cancer

Fig. 2

NSUN7 epigenetic loss abrogates m5C methylation of the CCDC9B mRNA leading to diminished RNA stability and protein downregulation. (A) Restoration of NSUN7 protein expression by stable transduction in epigenetically-silenced SNU-423 HCC cells, analyzed by western blot. (B) RNA bisulfite sequencing of multiple clones of CCDC9B transcripts from empty vector (EV) and NSUN7-transfected SNU-423 cells. Cytosines are represented as short vertical lines, the bsRNAseq target C1324 is indicated by a black arrow, and presence of a methyl group is represented as a black square. (C) RNA bisulfite sequencing of CCDC9B transcripts from the HCC cell panel plus a normal liver sample, according to the basal NSUN7-promoter methylation status in liver cancer cell lines. (D) Efficient CRISPR-Cas9 mediated knockout (KO) of NSUN7 protein expression in the hypomethylated and expressing HEP3B2-1-7 HCC cell line. (E) RNA bisulfite sequencing of the CCDC9B transcript from NSUN7 WT and NSUN7 KO HEP3B2-1-7 cells. (F) CCDC9B transcript half-life analyzed by Actinomycin D chase assay (left) and CCDC9B protein levels (right) in EV and NSUN7-transfected SNU-423 cells. (G) CCDC9B transcript half-life analyzed by Actinomycin D chase assay (left) and CCDC9B protein levels (right) in NSUN7 WT and KO HEP3B2-1-7 cells

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Molecular Cancer

ISSN: 1476-4598