Fig 1: MYC was an upstream regulator of miR-937-3p. A Binding motif of MYC in the promoter of miR-937-3p, predicted by JASPAR (http://jaspar.genereg.net/). B The MYC expression level in 40 LUAD tissues and paired adjacent tissues was investigated by qRT-PCR. C MYC was closely related with survival probability. D miR-937-3p gene promoter region. E Chromatin immunoprecipitation (ChIP) assay indicated the MYC binds to the putative binding site upstream of miR-937-3p. F, G The relative expression levels of miR-937-3p and SOX11 responding downregulated MYC were detected by qRT-PCR. H Levels of miR-937-3p and SOX11 was accessed by qRT-PCR after co-transfection. I, J Migration and invasion was determined after co-transfected; scale bars, 50 µm; scale bars, 200 µm. The data expressed as the mean ± SD. (*P < 0.05; **P < 0.01; ***P < 0.001)
Fig 2: miR-937-3p promotes metastasis via targeting SOX11. A, C Visualization of the entire lung and HE-stained lung sections after injecting transfected NSCLC cells into nude mice. B The number of lung metastatic sites was counted under the microscope. D SOX11 and MMP2 expression levels the samples collected from tumor nodules were analyzed by IHC; scale bars, 50 µm. The data expressed as the mean ± SD. (*P < 0.05; **P < 0.01; ***P < 0.001)
Fig 3: SOX11 is a direct downstream miR-937-3p target in LUAD cells. A Predicating candidate gene targets by intersecting from three different bioinformatics websites (TargetScan, miRDB and miRWalk). B The SOX11 expression level in 60 LUAD tissues and paired adjacent tissues was investigated by qRT-PCR. C The relationship between SOX11 and the survival of LUAD patients (n = 366 in the miR-937-3p-low group and n = 353 in the miR-937-3p-high group). D A negative regulatory effect of miR-937-3p on SOX11 was tested by qRT-PCR. E Predicted miR-937-3p seed region at the 3'- UTR of SOX11. F Luciferase activity was analyzed in LUAD cells co-transfected with miR-937-3p-mimics or negative control with pmirGLO-SOX11-Wt or pmirGLO-SOX11-Mut. G The expression levels of SOX11 protein and AKT pathway related proteins in the transfected A549 and H1395 cells were analyzed by Western blot. 1: NC;2: miR-937-3p;3: inhibitor NC;4: miR-937-3p inhibitor; GAPDH was used as a control. The data expressed as the mean ± SD. (*P < 0.05; **P < 0.01; ***P < 0.001)
Fig 4: SOX11 overexpression reversed the effect of miR-937-3p on the malignant progress of LUAD cells. A After co-transfection and the expression level of SOX11 was detected by qRT-PCR. B The roles of promoting angiogenesis of miR-937-3p was attenuated by high expression of SOX11; scale bars, 100 µm. C, D Transwell assays and wound-healing assay detected that SOX11 attenuated the inhibitory effects of miR-937-3p; scale bars, 200 µm. E, F The expression levels of SOX11 protein and AKT pathway related proteins after SOX11 plasmid transfection were analyzed by Western blotting in co-transfected cell lines. 1: miR-NC;2: miR-NC+SOX11;3: miR-937-3p;4: miR-937-3p+SOX11; The data expressed as the mean ± SD. (*P < 0.05;**P< 0.01; ***P < 0.001)
Fig 5: Schematic diagram of MYC/miR-937-3p/SOX11/PI3K/AKT pathway
Supplier Page from Abcam for Anti-SOX11 antibody [EPR20457]