Fig. 5: The feedback loop of ANKHD1/ MALAT1/YAP1 is responsible for YAP1 activity.

A Correlation of the expression of ANKHD1 and MALAT1 was analyzed in colorectal cancer using the GEPIA database (P < 0.05). B, C qRT-PCR was performed to detect the mRNA expression of ANKHD1, MALAT1, and YAP1 in ANKHD1-silenced or MALAT1-silenced cells, and the data are presented as the mean ± SD (**P < 0.01, ***P < 0.001). D Immunofluorescence staining indicated the colocalization of ANKHD1 and MALAT1 in the cytoplasm. E, H RNA-immunoprecipitation assay showed the binding of ANKHD1 or YAP1 with MALAT1. qRT-PCR was used to detect the RNA level of MALAT1 in the precipitates, and data are presented as the mean ± SD (**P < 0.01, ***P < 0.001). F RNA pull-down experiment showed the interaction between ANKHD1 or YAP1 and MALAT1, biotin-labeled MALAT1 was incubated with HCT116 cell lysates, and the enriched ANKHD1 or YAP1 was detected by Western blotting. The antisense MALAT1 was used as control. G Correlation of the expression of MALAT1 and YAP1 was analyzed in colorectal cancer using the GEPIA database (P < 0.05). I A nucleus-cytoplasm separation assay showed that MALAT1 silencing decreased YAP1 expression in the nucleus and increased the phosphorylation of YAP1 in the cytoplasm. J Western blot analysis showed that MALAT1 silencing increased the degradation of YAP1 after treatment with CHX (10 μg/ml). K MALAT1 silencing inhibited the expression of YAP1, as well as CTGF downstream of YAP1. Data are presented as the mean ± SD (**P < 0.01, ***P < 0.001).