title: GeoMx digital spatial profiling on enthesis biopsies from psoriatic arthritis patients before and after treatment with secukinumab
design: GeoMx digital spatial profiling on enthesis biopsies from psoriatic arthritis patients before and after treatment with secukinumab
library_construction_protocol: Entheseal biopsy of the extensor common tendon´s enthesis was performed. Briefly, biopsies were done by Blakesley forceps in 60° flexed position after identifying the lateral epicondyle, the olecranon and the radial head by ultrasound. Entheseal tissue slides from six patients before and after treatment with secukinumab were analyzed with GeoMX Digital Spatial Profiler (Nanostring) Tissue slides were prepared following the RNA Slide Preparation Protocol in the GeoMx NGS Slide Preparation User Manual (NanoString, MAN¬ 10150-06). Briefly, slides were baked at 60°C for 60 min and then treated sequentially with HIER (ER2, 20 min, 99°C) and 1 mg/mL proteinase K (Ambion, cat. 2546) in 1X phosphate-buffered saline (PBS) at 37°C for 15 min. After pretreatment, slides were hybridized with the GeoMx Whole Transcriptome Atlas Panel (GMX-RNA-NGS-HuWTA-4). The library preparation and purification were carried out using GeoMX DSP NGS Readout Protocol (MAN-10153-06).