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. 2021 Nov 3;12(11):1046. doi: 10.1038/s41419-021-04315-1

Fig. 5. DNA hypermethylation causes miR-497-5p silencing and suppresses choriocarcinoma CSLCs.

Fig. 5

A Profiling of methylation levels of CpG sites in the miR-497-5p upstream region is presented as an epigram (left) and average methylation level of miR-497-5p promoter between the indicated groups (right). B qRT-PCR demonstrated miR-497-5p expression in CSLCs after treatment with decitabine, zebularine, or 5-zazcytidine for 7 days compared with control groups. C Western blot analysis of SALL4 protein levels (up) and quantification (bottom) with zebularine treatment. D Western blot analysis of SALL4 protein levels (up) and quantification (bottom) with zebularine treatment and anti-miR-497-5p. E qRT-PCR demonstrated miR-497-5p expression in CSLCs after treatment with 80 μM zebularine and overexpressed SALL4 compared with control groups. F Representative images of sphere (left, magnifications: ×100) and sphere counts (right) with different groups. G Xenograft tumor in nude mice treated with zebularine at 200 mg/kg/day for 10 days. H Tumor volumes were measured every 4 days for 3 weeks (left). Tumor weight in the mice with zebularine treatment (middle). Survival curve of tumor-bearing mice between the zebularine group and control group (right). Data represent mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.