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. 2021 May 14;12(5):493. doi: 10.1038/s41419-021-03773-x

Fig. 4. miR-135b activates the NF-κB signaling.

Fig. 4

A, B Activity of the NF-κB luciferase reporter gene in 293FT (A) transfected with miR-135b mimics, miR-135b inhibitors and corresponding negative control (NC) and in A549 stable cell lines (B). C, D Real-time PCR analysis of the mRNA expression of NF-κB downstream genes in the indicated cells. E Western blotting detecting the protein level of NF-κB downstream genes, Bcl-2, Bcl-xL, Cyclin D1, and MMP9 in indicated cells. β-tubulin served as a loading control. *Non-specific band. F Western blotting detecting the protein level of NF-κB signaling, p65, phospho-p65 (p-p65), IKKβ, phospho-IKKα/β (p-IKKα/β), in the indicated cells stimulated with TNF-α (10 ng/ml) for 20 min. β-tubulin served as a loading control. G Representative images of immunofluorescence staining of p65 (labeled with TRITC) in the indicated cells stimulated with TNF-α (10 ng/ml) for 20 min. Scale bars, 20 μm. Error bars represent the mean ± SD obtained from three independent experiments. p values are calculated by a two-tailed, unpaired t-test. *p < 0.05; **p < 0.01; ***p < 0.001.