Gene Target Identification and Validation.

Publically available RNA-sequencing and microarray datasets were used to identify the top 100 genes highly expressed in HCC with very low to no expression in other tissues and blood components (15–17). The low expression of candidate genes within WBCs persisting in the CTC-iChip output was confirmed by RNA-sequencing of processed blood from healthy donors, and qRT-PCR of WBCs purified from whole blood was used as an additional exclusion criterion. Ten genes were selected to establish a signature of HCC-derived CTCs, enriched within a background of normal blood cells.