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Fig. 4 | Molecular Cancer

Fig. 4

From: Long noncoding RNA GAS5 inhibits progression of colorectal cancer by interacting with and triggering YAP phosphorylation and degradation and is negatively regulated by the m6A reader YTHDF3

Fig. 4

YTHDF3 is a novel target of YAP and can be regulated by GAS5. a The correlation of log2 fold change (FC) of differentially expressed genes in YAP knockdown cells and log2 (FC) of differentially expressed genes in GAS5 overexpressing cells are indicated. Genes were further stratified into groups based on the correlation between the YAP knockdown and GAS5 overexpression change. Group 1 (pink) were genes that were down-regulated in both treatment groups. Group 9 (purple) were those genes that were up-regulated in both treatment groups. (/log FC/ ≥ 1, p-value< 0.05 in RNA-seq). b Gene ontology (GO) analysis of differentially expressed genes (FC > 2; p-value < 0.05 in RNA-seq) in YAP knockdown and GAS5 up-regulation groups. c-d qRT-PCR detection of genes expressions of YAP target genes. All experiments were performed in triplicate, and results are presented as mean ± SD. **P < 0.01, and ***P < 0.001. e-f Analysis of the indicated protein expression in DLD1 cells transfected with YAP-specific siRNA or si-control (e), and HCT116 cells transfected with YAP plasmid or vector (f). g-h qRT-PCR and Western blots analysis showed a positively correlation between YAP and YTHDF3 in various CRC cells. i Binding of YAP to the YTHDF3 promoter was studied by chromatin immunoprecipitation (ChIP) assay. And the coprecipitated DNA was subjected for analysis of YTHDF3 by qRT-PCR (upper panel). The PCR procedures was shown by agarose gel electrophoresis (bottom panel). Experiments were performed in triplicate, and data are presented as mean ± SD. ***P < 0.001 (j) Potential YAP binding sites in the human YTHDF3 promoter between − 550 and + 100 bp. YAP plasmid or YAP-specific siRNA were transfected into HEK293 cells to detect the transcriptional activity of YTHDF3 promoter by dual luciferase reporter system. Experiments were performed in triplicate, and data are presented as mean ± SD. ***P< 0.001 (k-l) Analysis of the indicated protein expression in HCT116 and DLD1 cells co-transfected with GAS5 and YTHDF3 plasmid or YTHDF3 alone (k), and LOVO and RKO cells co-transfected with sh-GAS5 and sh-YTHDF3 or sh-GAS5 alone (l)

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