Fig 1: TMED6 is the putative target for aptamer 1-717.a Vulcano plot of aptamer 1-717 over m12-3773 binding on protein arrays Two sides paired T test was performed. b Cold target inhibition assay. cy3-labeled aptamer 1-717 was admixed at different molar ratios with unlabeled recombinant TMED6. The mixture was then used as a probe against a section of human pancreata. Sections were counterstained with anti-insulin antibody and DAPI; images were acquired and processed with Cell Profiler. Data derived from two independent experiments evaluating 450, 528, 449, and 491 INS+cells and 2590, 3063, 2875, and 2775 cells INS-cells for 0, 1, 3, and 10 rTMED6/aptamer ratio, respectively. The box plot shows the median, the 25, 75, 10, 90, 5, and 95th percentile. One-way ANOVA followed by Holm–Sidak multiple pairwise comparisons was performed. c Aptamer 1-717 was run at the indicated concentration as analyte against recombinant TMED6 (ligand) in a surface plasmon resonance assay. d Affinity. MIN6 cells were stained with vital dye, and different concentration 1-717 aptamer-chimera annealed with cy5-labeled guide saRNA and analyzed by flow cytometry. e Human islets (250 IEQ) were treated with siRNATMED6 aptamer m12-3773 chimera or with siRNAscrambled aptamer m12-3773 chimera and TMED6 expression was assessed by qRT-PCR 72 h later. Two sides paired T test was performed. f Part of the islets from (e) were dissociated and labeled with 1-717 aptamer and anti-insulin antibody and analyzed by flow cytometry. Two sides paired T test was performed. g Correlation between aptamer 1-717 signal and TMED6 expression. Source data are provided as a Source Data file.
Supplier Page from Abcam for Recombinant Human TMED6 protein