Quantitative Assessment of TRIM22 Expression

Deparaffinised and rehydrated tissue sections were processed for epitope retrieval using EnVision FLEX target retrieval solution (Agilent, K8004), as per manufacturer’s instructions. Samples were processed and stained for TRIM22 (Proteintech, 13744-1-AP; 1/50 dilution) and FLEX haematoxylin (Agilent, K8008) using a Autostainer Link 48 (Agilent), as per manufacturer’s instructions. TRIM22 antibody specificity was validated by staining sectioned MRC5 (positive control) or A549 (negative control) cell pellets. Tissue sections were independently assessed for TRIM22 expression by a qualified pathologist. Automated quantitation of TRIM22 expression levels was performed using whole-slide scans and Image-Pro Premier (Media Cybernetics), as previously described (Leeming et al., 2015 (link); Akram et al., 2018 (link)).