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Fig. 3 | Molecular Cancer

Fig. 3

From: The long noncoding RNA LINC00312 induces lung adenocarcinoma migration and vasculogenic mimicry through directly binding YBX1

Fig. 3

LINC00312 affected angiogenesis and VM formation via YBX1. a Fluorescent in situ hybridization analysis of LINC00312 subcellular localization. b LINC00312-sense and LINC00312-antisense probes were biotinylated, transcribed in vitro, and incubated with PC-9 whole-cell lysates for RNA pull-down assays. The silver staining represented the two LINC00312–specific bands (red arrows). Schematic repeatedly appeared in three independent assays. c LINC00312–specific bands were excised and analyzed by Western blot assay. d RIP assays were performed using antibodies against YBX1. Real-time PCR (RT–PCR) were normalized to the housekeeping gene GAPDH. Shown is one representative assay from at least three independent replicates. e Transwell migration assays of PC-9 cells transfected with the indicated plasmids. Bar charts showing depleting YBX1 block LINC00312-mediated metastasis. f Tube formation assay of VM in PC-9 cells transfected with the indicated plasmids. g LINC00312 regulates YBX1-related angiogenic genes of VE-cadherin, TGF-β, VEGF-A, VEGF-B and VEGF-C in PC-9. VE-cadherin, TGF-β, VEGF-A and VEGF-C expression increased markedly following overexpression of LINC00312. VE-cadherin, TGF-β, VEGF-A and VEGF-C decreased following knockdown of LINC00312. Control represent wild cells transfected with vector plasmid. Error bars represent SD from three reactions. Significance was calculated using Student’s t test. *p < 0.05; **p < 0.001; ***p 0.0001; ns, not significant. Scale bars: 100μm in 600 x images

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