Ish probe

The ISH probe used for detecting PTTG3P-labeled digoxin was designed and synthesized by Exiqon (Shanghai, China). ISH was performed using the ISH Kit (Boster Bio-Engineering Company, Wuhan, China) and the stained tissue sections were reviewed and scored separately by two pathologists blinded to the clinical parameters. The score standard for the staining intensity was as follows: 0 (negative), 1 (weak), 2(medium), and 3 (strong); and 0 (0%), 1 (1–25%), 2(26–50%), 3 (51–75%), and 4 (76–100%) for the staining extent. For statistical analysis, a final staining score of 3 or higher was considered to be high expression, respectively.

The ISH probe used for detecting lncRNA UBE2CP3-labeled digoxin was designed and synthesized by Exiqon (Shanghai, China). The probe sequence was designed as TGTGTCACTAGGCATTGT. ISH was performed using the ISH Kit (Boster Bio-Engineering Company, Wuhan, China) according to the manufacturer’s instructions. The ISH staining regions for lncRNA UBE2CP3 were scored by two pathologists, blinded to the clinical parameters. The score standard for the staining intensity was defined as follows: 0 (negative), 1 (weak), 2 (medium), 3 (strong). The score of staining extent was 0 (10%), 1 (11%–25%), 2 (26%–50%), 3 (51%–75%), and 4 (76%–100%). The final lncRNA UBE2CP3 expression score was calculated with the intensity score and extent score, ranging from 0 to 7. The high expression group (positive group) was defined as a total score of 3 or higher [24 (link)].