biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsYu-Shan ChenIllumina NovaSeq 6000Agilent Bioanalyzer 2100 (or TapeStation); Qubit 4 Fluorometer (Thermo Fisher Scientific)NanoDrop ND-1000 spectrophotometerRNA-seq of total RNAHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16607Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy characterised by rapid metastasis and marked chemoresistance. The absence of early symptoms and reliable diagnostic markers makes timely detection clinically challenging, and progress in developing new therapeutics has been limited. This project aims to establish a new class of non-toxic drug sensitiser, N-methyl-(5-pyrid-3-ylthien-2-yl)methylamine (B10), termed a binary drug (BD), identified through screening of a Maybridge chemical fragment library, which enhances gemcitabine-induced killing of pancreatic cancer cells when co-administered by targeting drug-resistance-related efflux transporters and key oncogenic pathways.biostudies-arrayexpressLibrary Construction - Stranded, polyA-enriched mRNA libraries were constructed from 1 µg total RNA using the Illumina TruSeq Stranded mRNA Library Prep Kit following the manufacturer's protocol.Sequencing - Libraries sequenced on Illumina NovaSeq 6000 using S1-100 flow cell in paired-end 75 bp mode at 36–45 million reads/sample (~200–300 pM loading). Dual-indexing for multiplexing; cluster generation via NovaSeq 500/6000 Reagent Kit.Sample Treatment - PANC-1 cells were seeded at 5,000 cells/well in 96-well plates, pre-treated with 10 µM BD B10 for 24 h, then the medium was replaced with fresh medium containing 0.1 µM gemcitabine + 10 µM BD B10 for an additional 96 h incubation at 37°C, 5% CO₂. Control conditions included: (1) untreated, (2) BD B10 alone (10 µM), and (3) gemcitabine alone (0.1 µM).Growth Protocol - PANC1 cells were cultured in DMEM with 10% heat-inactivated FBS and 1% penicillin-streptomycin at 37°C, 5% CO₂. Cells were passaged 3–4 days (1:3 ratio, 0.05% trypsin for 5 minutes at 37°C).Sample Collection - Treated PANC1 cells were harvested by trypsinisation (0.05% trypsin, 5 minutes at 37°C) at the endpoint of treatment (96 hours post-co-administration of gemcitabine and BD B10). Cells were pelleted by centrifugation (1000 g, 5 minutes, room temperature), washed three times with 1x PBS, and stored at -80°C.Nucleic Acid Extraction - Total RNA was extracted from snap-frozen Panc1 cell pellets using the RNeasy Plus Mini Kit (74136, Qiagen) according to the manufacturer's instructions. RNA quantity and purity were assessed using a NanoDrop ND-1000 spectrophotometer.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesYu-Shan ChenfalseRNA-seq analysis of co-treatment with gemcitabine and N-Methyl-(5-pyrid-3- ylthien-2-yl)methylamine in PANC-1 cellsPancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy characterised by rapid metastasis and marked chemoresistance. The absence of early symptoms and reliable diagnostic markers makes timely detection clinically challenging, and progress in developing new therapeutics has been limited. This project aims to establish a new class of non-toxic drug sensitiser, N-methyl-(5-pyrid-3-ylthien-2-yl)methylamine (B10), termed a binary drug (BD), identified through screening of a Maybridge chemical fragment library, which enhances gemcitabine-induced killing of pancreatic cancer cells when co-administered by targeting drug-resistance-related efflux transporters and key oncogenic pathways.2026-02-12T00:00:00Z2026-02-12T14:24:29.386Z2026-02-09T12:18:52.215ZE-MTAB-16607ERP188880EFO_0002944EFO_0004170EFO_0009653EFO_0003789EFO_0005518EFO_0004184EFO_0003969