Fig. 10

LINC00301’s oncogenic effects are also in part attributed to sponging miR-1276, and then triggering HIF1α. a Detection of targeted miRNAs using qPCR in the sample pulled down by biotinylated LINC00301 (sense and antisense). b Left: Sequence alignment of miR-1276 with the presumed binding sites within the WT or MUT LINC00301 regions. The luciferase report assay proved that miR-1276 overexpression could decrease the intensity of fluorescence in A549 (middle) and SPC-A-1 (right) cells transfected with the LINC00301-WT vector, whereas it had no impact on the LINC00301-MUT vector. c The 3′-UTR of HIF1A harbors 2 miR-1276 equivalent sites. Relative luciferase activity of reporter vectors containing HIF1A WT or MUT 3′UTR in A549 and SPC-1-A cells co-transfected with miR-NC or miR-1276. d The relationship between HIF1A mRNA expression and miR-1276 expression in NSCLC tissues. e The role of miR-1276 on HIF1A mRNA expression in A549 and SPC-A-1 cells. f The role of miR-1276 on HIF1α protein expression in A549 and SPC-A-1 cells. g, h The role of pLenti-CMV-LINC00301, miR-1276, pLenti-CMV-HIF1A, or their interaction roles on HIF1α protein expression in A549 and SPC-A-1 cells. i Trypan blue staining tested cell viability in pLenti-CMV-LINC00301, miR-1276, pLenti-CMV-HIF1A, or their interaction transfected in A549 and SPC-A-1 cells. j Colony formation assays tested cell proliferation (seeded at 6-well plate) in pLenti-CMV-LINC00301, miR-1276, pLenti-CMV-HIF1A, or their interaction transfected in A549 and SPC-A-1 cells. k Transwell migration assay tested migration activity in pLenti-CMV-LINC00301, miR-1276, pLenti-CMV-HIF1A or their interaction transfected in A549 and SPC-A-1 cells. l Transwell invasion assay tested invasive activity in pLenti-CMV-LINC00301, miR-1276, pLenti-CMV-HIF1A, or their interaction transfected in A549 and SPC-A-1 cells. Assays were conducted in triplicate. *p < 0.05, means ± SD was shown. Statistical analysis was performed by Student’s t-test