Supplementary Figure S2 from Analytic Validation of RNA In Situ Hybridization (RISH) for AR and AR-V7 Expression in Human Prostate Cancer
Year: 2023
Type: supplementary-materials
Abstract: Supplementary Figure S2: AR-E1 RISH signals show greater nuclear localization compared to other genes and are decreased with RNase pre-treatment of slides prior to RISH assay. (A) AR-E1 RISH signal shows dramatic nuclear localization (arrows) with some cytoplasmic signal (arrowheads) across benign and malignant primary prostate tumors, including some that have been treated with androgen deprivation therapy. In contrast, other genes such as positive control gene PPIB (cyclophilin B) show only cytoplasmic signal. (B) Both intranuclear and cytoplasmic signals are markedly decreased and nearly undetectable after RNase, confirming both signals correspond to RNA. (C) qRT-PCR analysis of LNCaP and LAPC4 cells performed on nuclear and cytosolic fractions of RNA (accompanying immunoblot confirms appropriate laminin and GAPDH enrichment in each fraction) reveals abundance of unspliced AR RNA in the nuclear compartment of both cell lines, while spliced AR message is present in both compartments.
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4362400513