Fig. 4

LncGHRLOS was identified as a downstream target of ZCCHC4. (A) Volcano map can be used to visualize the overall distribution of differentially expressed genes. Two levels of difference multiple (log2(Fold change)) and significance level were evaluated. (B) Gene expression cluster analysis is used to judge the clustering pattern of gene expression in different treatments. (C) The protein sequences of all genes and found differentially expressed genes (gene sequences) were compared with Uniport database protein sequences, and then the comparison results were annotated by GO function according to the known protein GO annotation in Uniport database, and the pathway significance was tested using hypergeometric distribution. The GO Terms screening condition for significant enrichment was the p value of hypergeometric distribution test less than 0.05. (D) The identified differentially expressed genes were enriched by KEGG metabolic pathway using the clusterpro digestion package in R language. (F) Filtering process of target Genes among MeRIP-seq, RNA-Seq and literature search. (G-J) CLEC18B, Col5A3, CPA4 and LncGHRLOS mRNA levels were detected in 10 CRC and adjacent tissue samples (K) LncGHRLOS mRNA levels in 50 CRC cancer tissues and adjacent tissues were detected by qRT-PCR. (L) ZCCHC4 and LncGHRLOS were verified in 50 continuous CRC cases correlation between endogenous LncGHRLOS mRNA levels (M) Detection of LncGHRLOS mRNA levels after endogenous LncGHRLOS using NC or ZCCHC4 by lentivirus in different CRC cell lines. (N) RNA pulldown of endogenous LncGHRLOS using NC or ZCCHC4 probe. (O) RIP-qPCR assay of ZCCHC4 enrichment by LncGHRLOS in HCT-15 cell lines. (P) RNA stability of LncGHRLOS mRNA after treatment with ZCCHC4 overexpression and knockdown