Fig. 5

XIST indirectly regulated the VEGFA expression by acting as a ceRNA for miR-302b-3p. A Subcellular localization of XIST detected by nuclear and cytoplasmic RNA fractionation analysis and lncRNA FISH. GAPDH and U6 were used as nuclei and cytoplasmic references, respectively. Green fluorescence represented cyc3-labeled XIST probes and blue fluorescence represented DAPI-stained nuclei. B The expression of miRNAs (miR-302a-3p, miR-302b-3p, and miR-302d-3p) was evaluated by RT-qPCR in HLF and non-HLF cells. C The expression of miRNAs (miR-302a-3p, miR-302b-3p, and miR-302d-3p) was evaluated by RT-qPCR in HLF cells infected with the indicated adenovirus. D RNA pull-down was executed in HLF cells, followed by RT-qPCR to detect the enrichment of XIST. E The relative luciferase activities were measured in 293 T cells after transfection with XIST-WT or XIST-Mut and miR-302b-3p mimics or NC mimics, respectively. F The expression levels of VEGFA and miR-302b-3p were evaluated by RT-qPCR in HLF cells infected with the indicated adenovirus. G RNA pull-down was performed in HLF cells, followed by RT-qPCR to detect the enrichment of VEGFA. H Relative luciferase activity was measured in 293 T cells after transfection with VEGFA-WT or VEGFA-Mut and miR-302b-3p or NC mimics, respectively. I The mRNA expression of VEGFA in HLF cells infected with the indicated adenovirus was evaluated by RT-qPCR. J The expression levels of miR-302b-3p and VEGFA in 25 HLF tissues and 16 control LF tissues were detected by RT-qPCR, and VEGFA expression obviously upregulated in HLF tissues and miR-302b-3p expression markedly downregulated in HLF tissues. K Pearson correlation coefficient was used to analyze the correlation between the expression levels of XIST, miR-302b-3p, and VEGFA in HLF tissues. shNC, corresponding negative control of shXIST; shXIST, the RNAi adenovirus against XIST; OE-NC, corresponding negative control of OE-XIST; OE-XIST, the overexpression adenovirus of XIST; OE-miR-NC, corresponding negative control of OE-miR-302b-3p; OE-miR-302b-3p, the overexpression adenovirus of miR-302b-3p; sh-miR-NC, corresponding negative control of sh-miR-302b-3p; sh-miR-302b-3p, the RNAi adenovirus against miR-302b-3p. ^*P < 0.05, ^**P < 0.01, and.^***P < 0.001