Fig. 2

Screening and identification of lncRNA SPOCD1-AS. a The schematic protocol of lncRNA expression analysis in MeT-5A cells treated with EVs. b The heat map of differentially expressed lncRNAs (cancer EVs vs Ctrl, fold change > 1.5 or < 0.5, p < 0.05). The red arrow indicates lncRNA SPOCD1-AS. c qRT-PCR analysis of differentially expressed lncRNAs in MeT-5A cells treated with cancer EVs or Ctrl group. Data were normalized to GAPDH and presented as 2^-ΔΔCT. d Schematic annotation of SPOCD1-AS genomic locus on chromosome 1:31,789,130-31,791,322 in human. The black rectangles indicate exons. e UP: Agarose gel and sequencing of second-round products of 5′ RACE and 3′ RACE. The red arrows indicate the main PCR products. The vertical line indicates a predicted transcriptional start site or end site. The black arrows indicate transcriptional directions. Down: the full-length nucleotide sequence of SPOCD1-AS. The additional sequence according to Ensembl database reference sequence is marked red. f Northern blot using DIG-labeled probes and qRT-PCR analysis of SPOCD1-AS in IOSE-80 and two ovarian cancer cells. GAPDH was used as the loading control. qRT-PCR data were normalized to GAPDH and presented as 2^-ΔΔCT compared with IOSE-80 cells. g qRT-PCR analysis of SPOCD1-AS in EVs secreted from the three cell lines. Data were normalized to GAPDH and presented as 2^-ΔΔCT compared with IOSE-80 EVs. Data are representative of at least three independent experiments and are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001