Figure 2

Effects of miR-26a on toll-like receptor (TLR)3 signaling by gain or loss of miR-26a function in NR8383. (A) Stem loop RT-qPCR results of miR-26a expression, (B) RT-qPCR results of tlr3 mRNA expression, and (C) western blotting of TLR3 protein expression after 10nM miR-26a mimics or inhibitor was transfected into NR8383 macrophages for 48 h. (D) Western blotting results of TLR3 protein expression after 0.1, 1.0 and 10.0 nM of miR-26a mimics were transfected into NR8383 cells for 48 h. (E) Western blotting of TLR3 protein expression and (F) RT-qPCR results of ifn-β and tnf-α mRNA expression in NR8383 and ELISA results of TNF-α protein expression in cell supernatant (G) after incubated with 10 nM mimics and inhibitors for 24 h and poly I:C (PIC, TLR3 ligand, 10 μg/ml) stimulation for another 24 h. U6 snRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as internal controls in RT-qPCR for miRNA and mRNA expression detection respectively. Bars represent the standard error of the mean from three cell experiments. NC, negative control. One representative plot and quantitative data from three independent western blotting experiments are shown. Ratio indicates the optical intensity of TLR3 protein bands against GAPDH. *Statistically significant differences (P <0.05) against the NC; ^significant differences against the mock group (Mann–Whitney U-test).