Figure 2.

miR-155 directly inhibited the expression of BRG1 and up-regulated the subsequent VEGFC, but not VEGFD expression in NKTCL cells. A. The 3ʹ-UTR sequences of human BRG1 gene containing the wild-type (WT) or a mutated sequence (MUT) for the miR-155-binding site was cloned into the luciferase reporter construct and transfected into SNK-6 (left panel) or YTS (right panel) cells. The cells were then treated with either miR-155 mimics or negative control (NC), and the luciferase activity was measured. B. SNK-6 or YTS cells were transfected with either miR-155 inhibitor or NC. The expression of miR-155, BRG1, VEGFC, and VEGFD on the steady-state mRNA level was examined by RT-qPCR and presented as a ratio relative to that of the internal control (GAPDH). C. and D. The expression of BRG1, VEGFC, and VEGFD on the protein level from indicated cells was examined by Western blotting. The representative Western blotting image was shown in C and the quantification of relative protein levels to that of the internal control (GAPDH) shown in D. E. The secretion of VEGFC into the conditioned medium (CM) from SNK-6 or YTS cells upon treatment with miR-155 inhibitor or NC was measured by ELISA. The relative VEGFC level in CM from NC-treated cells was arbitrally defined as 1. F. and G. The expression of p-STAT3 and STAT3 from indicated cells was examined by Western bloting. The representative Western bloting image was shown in F and the quantification of relative protein levels to that of the internal control (GAPDH) shown in G. **P < 0.01; ***P < 0.001.