Fig. 5: Upregulation of FOXD2-AS1 bound to STAT3 and augmented STAT3 transcriptional activity by recruiting PRMT5.
a Distribution of FOXD2-AS1 was predicted by public webserver lncATLAS. b The expression levels of FOXD2-AS1 were determined by qRT-PCR in cytoplasmic and nuclear fractions of laryngeal cancer cells. The percentages of FOXD2-AS1 distribution in cytoplasm and nucleus were shown. c Real-time PCR analysis of FOXD2 was performed, showing that FOXD2-AS1 would not affect the expression of its host gene. d REPOTM STAT3 luciferase assay to assess STAT3 activation in the indicated cells transfected with STAT3 luciferase reporter plasmid, showing an increased activity of STAT3 in FOXD2-AS1-overexpressing cells. e Real-time PCR analysis of STAT3 target genes (MYC, BIRC5, and MCL1) mRNA expression in FOXD2-AS1-overexpressing and -silencing cells. Expression levels were normalized to GAPDH. f Western blot analysis of STAT3, p-STAT3 expression in indicated cells. Alpha-tubulin was used as a loading control. g, h RNA pull-down assay and co-immunoprecipitation (Co-IP) assay were performed to assess the relationship between FOXD2-AS1, STAT3, and PRMT5. g SDS–PAGE analysis of eluted proteins after RNA pull-down, showing that both STAT3 and PRMT5 were presented in the FOXD2-AS1 pull-down fraction. h Co-IP of STAT3 and PRMT5 was performed in FOXD2-AS1-silencing and control cells, showing that FOXD2-AS1 depletion disrupted the combination of STAT3 and PRMT5. *P < 0.05, **P < 0.01, and ***P < 0.001.
