Fig. 2

Mechanism of lncRNA methylation in GCSCs. A Tumorsphere formation assay of a single cell from the GCSCs sorted from HGC-27 (GCSC-HGC) and MGC-803 (GCSC-MGC). B Western blot analysis of the stemness genes in GCSC-HGC and GCSC-MGC. C Migration capacity of GCSCs (GCSC-HGC and GCSC-MGC). GCSC-HGC, GCNSC-HGC, GCSC-MGC, and GCNSC-MGC were analyzed with Transwell migration assays to examine the cell migration. The representative images of the migrated cells are indicated on the left. The percentage of the migrated cells is shown on the right (**, p < 0.01). Scale bar, 50 μm. D Efficiency of chemoresistance of GCSCs (GCSC-HGC and GCSC-MGC). GCSCs or GCNSCs were treated with cisplatin at different concentrations. At 48 h after treatment, the cells were characterized using cell viability assays (**, p < 0.01). The half maximal inhibitory concentration (IC₅₀) values were calculated. E Examination of the expressions of methyltransferases METTL3 and METTL14 in GCSCs. F Western blot for METTL3 in GCSCs with METTL3-shRNA or METTL3-shRNA rescue. G m6A level of lncRNAs in GCSC with METTL3-shRNA or METTL3-shRNA rescue. H Detection of the expressions of stemness genes in the METTL3-silenced or rescued GCSCs using Western blot. β-tubulin was used as a control. I Western blot for METTL14 in GCSCs with METTL14-shRNA or METTL14-shRNA rescue. J m6A level of lncRNAs in GCSC with METTL14-shRNA or METTL14-shRNA rescue. K Detection of the expressions of stemness genes in METTL14-silenced or rescued GCSCs by Western blot. **, p < 0.01