Fig. 8

LIF is directly targeted by miR-637 and circSEPT9 stimulates LIF-STAT3 pathway by sponging miR-637 in TNBC. a and b The relative expression of LIF were detected after transfection with miR-637 and miR-637 inhibitor with qRT-PCR and western blot. c Schematic illustration of LIF 3’UTR wide type (WT) and mutant (Mut) luciferase reporter vectors were shown. d The luciferase reporter assays showed that miR-637 directly bind to the 3′-UTR of LIF and inhibit luciferase activity. e and f The relative expression of LIF was assessed by IHC (magnification, × 200, Scale bar, 100 μm) and IF (magnification, × 100, Scale bar, 100 μm) assays. g The relative expression of LIF was determined in 60 paired TNBC tissues and adjacent normal tissues with qRT-PCR. h and i Pearson correlation analyses of LIF expression with miR-637 or circSEPT9 were shown in 60 TNBC tissues. j The relative expression of LIF in TNBC cells after transfection with circSEPT9 expression vector and si-circSEPT9 by qRT-PCR. k The relative expression of LIF after transfection or co-transfection with indicated vectors, siRNAs, miRNA mimics or inhibitors by qRT-PCR. l IF analysis was applied to detect the protein expression of LIF in TNBC cells transfected or co-transfected with indicated vectors, siRNAs, miRNA mimics or inhibitors (magnification, × 200, Scale bar, 50 μm). m The relative protein levels of LIF and downstream LIF-STAT3 pathway-related molecules were measured in TNBC cells after transfection or co-transfection with indicated vectors, siRNAs, miRNA mimics or inhibitors by western blot. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001