The top five downregulated genes in CHD6 KD group were PRRG4, SPAST, TMEM65, PLEKHA8, and ZBTB18. To determine these potential downstream targets that may mediate the oncogenic effect of CHD6 in CRC, we performed correlation analysis with TCGA dataset and found that TMEM65 mRNA expression was significantly associated with CHD6 (Supplementary Fig. S5a); moreover, survival analyses showed that high transcriptional activation of TMEM65 (Supplementary Fig. S5b), but not other four genes, was associated with worse prognosis for overall survival in CRC patients. This evidence concerns the gene SPAST and colorectal carcinoma.