RNA in situ Hybridization of LINC01187

RNA in situ hybridization (RNA-ISH) was performed using the RNAscope 2.5 HD Brown assay (Advanced Cell Diagnostics, Newark, CA) and target probes against LINC01187 (catalog no. 532311) as previously described (10 (link)). Dual RNA-ISH was performed using the RNAscope 2.5 HD duplex assay and target probes against L1CAM, LINC01187, and FOXI1 (catalog no. 567451,532311-C2, and 476359-C2). RNA quality was evaluated using positive control probe targeting human housekeeping gene PPIB. All evaluated cases passed RNA-ISH quality control (QC) except one ESC-RCC case. Assay background was monitored using a negative control probe targeting bacterial DapB gene. Stained slides were examined under 100x and 200x magnification for RNA signals in tumor cells and adjacent benign kidney tissues. The RNA-ISH assay stained each RNA molecule as an individual brown, punctate dot. The staining was independently assessed by three study participants including two pathologists (XWang, RMannan, and RMehra) at 100x, 200x and 400x magnification to assess for presence and patterns of expression.