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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue <t>microarrays.</t> G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.
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Image Search Results


A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue microarrays. G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.

Journal: Cell Death & Disease

Article Title: Multi-omics analysis reveals that ALYREF-mediated m 5 C modification promotes platinum resistance in ovarian cancer via the NSUN2/ALYREF/LGR4 axis

doi: 10.1038/s41419-025-08310-8

Figure Lengend Snippet: A Schematic diagram of the ALYREF m 5 C methylation binding site mutation. B m 5 C-RIP and qRT-PCR assays detecting the influence of ALYREF on the m 5 C methylation level of LGR4 mRNA. C qPCR detection of LGR4 RNA expression in the influence of ALYREF. D Western blot analysis of LGR4 protein expression in ALYREF-overexpressing A2780/DDP and SKOV3/DDP cells, and in cells with mutated ALYREF binding sites. E Actinomycin D assay assessing the impact of ALYREF on LGR4 mRNA stability. F Representative images of IHC staining of LGR4 expression levels in tissue microarrays. G Correlation analysis between ALYREF and LGR4 protein expression levels. H Schematic representation of the subcutaneous tumor model in cisplatin-resistant ovarian cancer-bearing mice by Generic Diagramming Platform (GDP) . I Final diameter and weight of the subcutaneous tumors. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001. ****: p < 0.0001.

Article Snippet: Following a rinse with PBS, the tissue microarrays were incubated at 4 °C for 16 h with anti-ALYREF antibody (16690-1-AP; Proteintech, China) and anti-LGR4 antibody (20150-1-AP; Proteintech, China), each at a 1:300 dilution.

Techniques: Methylation, Binding Assay, Mutagenesis, Quantitative RT-PCR, RNA Expression, Western Blot, Expressing, Immunohistochemistry