Fig. 6

The lncRNA NEAT1 upregulates STAT3 to promote the EMT in osteosarcoma cells by inhibiting miR-483. U2OS cells were transfected with a negative control (OE-NC) or NEAT1 overexpression plasmid (OE-NEAT1). NEAT1-overexpressing U2OS cells were then cultured in the absence or presence of 15 µM Stattic (STAT3 inhibitor). a, b The migration of U2OS cells receiving different treatments was examined using a scratch wound healing experiment (a) or transwell migration experiment (b). Scale bar: 200 μm. c The invasion of U2OS cells subjected to the indicated treatments was assessed using the transwell invasion experiment. Scale bar: 200 μm. d The relative expression of STAT3, N-cadherin, E-cadherin, Vimentin, and Snail in U2OS cells was determined using qRT-PCR and normalized to the GAPDH mRNA. e The phosphorylation of STAT3 and the levels of STAT3, STAT1, N-cadherin, E-cadherin, Vimentin, and Snail in U2OS cells were examined using western blotting. GAPDH was employed as the input control. All experiments described in this study were performed at least three times, which referred to biological replicates. Each biological replicate consisted of 3 wells with cells of the same passage, which referred to technical replicates. Error bars denoted mean ± SD. p values were calculated using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. ***p < 0.001, **p < 0.01, and *p < 0.05