Fig. 6.

Autophagy-reduced METTL3 upregulates LINC01615 in a m⁶A modification-dependent manner. A A dot blot assay was used to analyze the m⁶A content of total RNA in DLD1 and LoVo cells cultured without serum supplementation for different times. Methylene blue staining was used as a loading control. B qRT–PCR analysis of LINC01615 expression in DLD1 and LoVo cells cultured without serum supplementation for different times. C qRT–PCR analysis of LINC01615 expression in DLD1 cells transfected with METTL3 siRNA or control siRNA. D Representative images of LINC01615, METTL3 and G6PD analyzed by FISH or IHC in LINC01615 high-expression and LINC01615 low-expression tumors. E MeRIP-qPCR showing the m⁶A modification enrichment of LINC01615 in DLD1 cells after METTL3 depletion. F qRT–PCR analysis of the stability of LINC01615 in DLD1 cells with or without METTL3 deletion. G WB analysis of METTL3 protein stability in DLD1 cells cultured with or without serum supplementation for different times. H Representative confocal microscopy images of LC3B puncta distribution in DLD1 cells under normal or serum-free starvation conditions and treated with or without Baf-A1 (1000 × magnification). I Analysis of the expression of METTL3 and LINC01615 in DLD1 cells transfected with ATG5 siRNA, METTL3 overexpression plasmid, control vector or treated with 3-MA 30 min before the end of each experimental time. J WB analysis of METTL3 expression in DLD1 cells treated with or without Baf-A1 under serum-free starvation conditions. K Co-IP analysis was performed with specific antibodies against METTL3 and LC3B in DLD1 cells under normal or serum-free starvation conditions. WB analysis of the interaction between METTL3 and LC3B. Data are the means ± SDs (n = 3 independent experiments), *p < 0.05, **p < 0.01