Fig. 5

GATA3-AS functions as a guide lncRNA that targetedly promotes the m6A modification of GATA3 pre-mRNA. a Genomic organization of GATA3 and GATA3-AS transcription units. Vertical bars represent exons. Arrows display the direction of transcription. b and c RNA distribution analysis of GATA3-AS in subcellular fractions of SK-Hep1 and HCCLM3 cells assessed by qPCR. U6 served as the nuclear marker, β-actin served as cytoplasmic marker. d Western blot analysis of GATA3 expression in SK-Hep1 and HCCLM3 cells transfected with GATA3-AS siRNAs or the control. e and f RIP-seq of the enrichment of GATA3-AS on KIAA1429 relative to IgG. g Western blot analysis of KIAA1429 in protein samples pulled down by even and odd sets for GATA3-AS, and control LacZ probes pool in SK-Hep1 and HCCLM3 cells. h and i ChIRP-qPCR analysis of the enrichment of GATA3-AS and GATA3 pre-mRNA in both even and odd sets relative to control LacZ probes pool in SK-Hep1 and HCCLM3 cells. j and k RIP-qPCR analysis of the enrichment of GATA3 pre-mRNA on KIAA1429 relative to IgG in control and GATA3-AS-depleted cells. l and m MeRIP-qPCR analysis of GATA3 pre-mRNA in SK-Hep1 and HCCLM3 cells transfected with GATA3-AS siRNAs or the control. n and o RIP-qPCR analysis of the enrichment of GATA3 pre-mRNA on HuR relative to IgG in control and GATA3-AS-depleted cells. p Relative luciferase activity in HEK293T cells co-transfected with luciferase reporter pmirGLO-GATA3-WT or pmirGLO-GATA3-MUT and GATA3-AS siRNAs or the control. Data are shown as the relative ratio of firefly luciferase activity to renilla luciferase activity. q Western blot analysis of GATA3 expression in SK-Hep1 and HCCLM3 cells co-transfected with GATA3 CDS-3′ UTR or GATA3 CDS vector and KIAA1429 siRNAs, GATA3-AS siRNAs or the control. Data are presented as mean ± SEM. NS: not significant; **P < 0.01, ***P < 0.001