anti alyref  (Proteintech)

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 Peak distribution of m 5 C modifications in A2780/DDP and parental cells. B Overall differential distribution of m 5 C modifications, illustrated as a pie chart, in A2780/DDP and parental cells. C Sequence context analysis of the m 5 C sites, displaying the nucleotide probabilities for 10 bases upstream and downstream. D RNA-Seq analysis of ALYREF knockdown A2780/DDP cells and schematic representation of overlapping genes identified by m 5 C-BIS-Seq in A2780/DDP and parental cells. E KEGG and GO enrichment analysis of genes identified in both RNA-Seq and m 5 C-BIS-Seq datasets. F Intersection of RIP-Seq data with genes positively regulating the Wnt signaling pathway, depicted schematically. G qPCR detection of LGR4, SPIN1, RBPJ, USP34, KANK1, DIXDC1, DAB2, and WNK2 expression in A2780/DDP and SKOV3/DDP ovarian cancer cells. 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: Sequencing, RNA Sequencing, Knockdown, Expressing

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 IGV visualization of m 5 C enrichment peak changes in the CDS, StopC and StartC regions of LGR4 mRNA in A2780/DDP and parental cells. B IGV visualization of LGR4 enrichment peaks in the Input group and ALYREF-IP group. C Western blot analysis of LGR4 protein expression in ovarian cancer cells with ALYREF knockdown and overexpression. D Immunofluorescence assay to assess the impact of ALYREF knockdown on LGR4 protein levels in A2780/DDP and SKOV3/DDP cells. E Subcellular distribution of LGR4 RNA in the cytoplasm and nucleus. F Actinomycin D assay assessing the impact of ALYREF knockdown on LGR4 mRNA stability. Data are displayed as mean ± SD; *: p < 0.05. **: p < 0.01. ***: p < 0.001.

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: Western Blot, Expressing, Knockdown, Over Expression, Immunofluorescence

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

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 This schematic illustrates that NSUN2-mediated m⁵C modification of LGR4 mRNA is recognized and bound by ALYREF, which enhances the stability and nuclear export of LGR4 transcripts. B Increased LGR4 protein expression subsequently activates β-catenin signaling, resulting in β-catenin accumulation and downstream activation of the Wnt/β-catenin pathway. C Activation of this signaling cascade ultimately leads to enhanced cisplatin resistance in ovarian cancer. Solid arrows represent activation events, dotted arrows indicate potential intermediate processes, and yellow arrows denote upregulated molecular changes.

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: Modification, Expressing, Activation Assay