Journal: Scientific Reports

Article Title: Role of stromal activin A in human pancreatic cancer and metastasis in mice

doi: 10.1038/s41598-021-87213-y

Figure Lengend Snippet: Activin A levels are significantly higher in tumor tissue and correlate to worse prognosis in PDAC. ( A ) Representative cores of pancreatic tumors from a human pancreatic tissue microarray stained with H&E to assess the percentage of epithelial versus stromal cells in each core. Adjacent sections were immunostained for activin A expression. ( B ) Quantification of the percentage of epithelial cells (left panel) or stromal cells (right panel) in normal tissue (white bar) versus tumor tissue (black bar). Statistical analysis is unpaired t-test. ( C ) Quantification of activin A in TMA (n = 63). An average of the percentage of epithelial and stromal fraction was calculated and scored in each TMA. Comparing epithelial (left panel) and stromal fraction (right panel) in normal tissue (white bar) versus tumor tissue (black bar). Statistical analysis 2-way ANOVA; mean ± SEM, ***(p < 0.001). ( D ) Kaplan–Meier plots of patient overall survival versus expression of activin A in either epithelial cells (left panel) or stromal cells (right panel). Statistical analysis Log-rank (Mantel-Cox) test, ** (p < 0.01), ***(p < 0.001).

Article Snippet: PDAC tissue microarray was commercially purchased from US Biomax Inc.

Techniques: Microarray, Staining, Expressing

Journal: Cell Death Discovery

Article Title: ALYREF-JunD-SLC7A5 axis promotes pancreatic ductal adenocarcinoma progression through epitranscriptome-metabolism reprogramming and immune evasion

doi: 10.1038/s41420-024-01862-2

Figure Lengend Snippet: A The mRNA expression profiles of m 5 C regulators in PDAC and normal tissues were based on TCGA+GTEx databases. B The mRNA expression of ALYREF in PDAC and adjacent tissues were obtained from GSE15471 and GSE16515 datasets. C ALYREF protein levels were analyzed in PDAC and paired normal tissues by western blotting (n = 9). D Representative IHC images and IHC scores of ALYREF staining in 20 pairs PDAC and normal tissues (scale bar, 250 μm (100×), 50 μm (400 ×)). E , F mIHC was performed on a tissue microarray to detect the relationship between ALYREF and CD8 + T cells ( n = 135, scale bar, 50 μm). G Kaplan–Meier curve of overall survival was performed based on the follow-up data of TMA assay. H Multivariate analysis of several factors was performed in TMA assay. Data are presented the mean ± SD. **** p < 0.0001.

Article Snippet: The human PDAC tissue microarray was created by Wuhan Servicebio technology (Wuhan, China) using 156 PDAC tissue specimens from the First Affiliated Hospital, School of Medicine, Zhejiang University, China.

Techniques: Expressing, Western Blot, Staining, Microarray

Journal: Cell Death Discovery

Article Title: ALYREF-JunD-SLC7A5 axis promotes pancreatic ductal adenocarcinoma progression through epitranscriptome-metabolism reprogramming and immune evasion

doi: 10.1038/s41420-024-01862-2

Figure Lengend Snippet: A Venn Diagram showing the intersection of RIP-BisSeq, RNA-seq and JASPAR database. JunD is identified as a potential target. B , C Expression of JunD following ALYREF knockdown was detected by RT-qPCR and western blotting. D RIP assay was used to validate the direct binding between ALYREF and JunD. E The mRNA decay rate of JunD in shNC and shALYREF PDAC cells treated with Actinomycin D. F The dot blot assay was used to assess the change of mRNA m 5 C level after NSUN2 knockdown in PDAC cells. G MeRIP-qPCR assay was conducted using m 5 C-specific antibody to measure the m 5 C levels. H RIP-qPCR assay was performed using anti-ALYREF antibody to determine the binging efficiency between ALYREF and JunD mRNA. I , J Expression of SLC7A5 following JunD knockdown was detected by RT-qPCR and western blotting. K Schematic illustration showing the position of ChIP- qPCR primers. L , M ChIP assay was performed to test whether JunD could bind to the promoter of SLC7A5. Dual-luciferase assays verified that whether JunD could promote the transcription of SLC7A5 mRNA. Data are presented the mean ± SD of 3 independent experiments, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The human PDAC tissue microarray was created by Wuhan Servicebio technology (Wuhan, China) using 156 PDAC tissue specimens from the First Affiliated Hospital, School of Medicine, Zhejiang University, China.

Techniques: RNA Sequencing, Expressing, Knockdown, Quantitative RT-PCR, Western Blot, Binding Assay, Dot Blot, ChIP-qPCR, Luciferase

Journal: Cell Death Discovery

Article Title: ALYREF-JunD-SLC7A5 axis promotes pancreatic ductal adenocarcinoma progression through epitranscriptome-metabolism reprogramming and immune evasion

doi: 10.1038/s41420-024-01862-2

Figure Lengend Snippet: A The expression levels of ALYREF, JunD and SLC7A5 in TMA were measured by multiplex immunohistochemical and two groups of representative multiplexes immunohistochemical images are shown (scale bar, 50 μm). B The correlation of among the expression levels of ALYREF, JunD and SLC7A5 in TMA samples was analyzed by Pearson correlation coefficient ( n = 139). C Overall survival analysis based on TMA cohort showed that patients with high co-expression of ALYREF, JunD and SLC7A5 have poor prognosis. D The graphic illustration of ALYREF modulating tumor progression and immune escape in PDAC.

Article Snippet: The human PDAC tissue microarray was created by Wuhan Servicebio technology (Wuhan, China) using 156 PDAC tissue specimens from the First Affiliated Hospital, School of Medicine, Zhejiang University, China.

Techniques: Expressing, Multiplex Assay, Immunohistochemical staining