For OCT samples, adaptations recommended by 10x Genomics were applied to preserve nuclear integrity. Filter sizes (30–40 µm) and wash stringency were optimized empirically to maximize nuclei recovery while minimizing debris. Single-nucleus libraries were prepared using the 10x Genomics Chromium Fixed RNA Profiling platform following manufacturer instructions (Pleasanton, CA, USA). Input targets were approximately 2,000–10,000 nuclei per sample where possible, with a sequencing depth target of ~50,000 reads per nucleus. Libraries were sequenced on Illumina platforms to a combined depth of approximately 700 million reads across experiments. Samples from distinct subjects were multiplexed (pooled and indexed with unique barcodes) and ran as a single run.