Fig. 8

miR-26a and miR-136 target TLR3 and RIG-I, respectively. a and b Graphical illustrations of the conserved miR-26a- and miR-136-binding motifs at the 3′-UTR of TLR3 and RIG-I, respectively. c and d Luciferase function was evaluated with luciferase reporter products comprising either WT or mutated (MU) copy of TLR3 and RIG-I after transfection with miR-26a/miR-136 mimic. Luciferase function was normalized to β-galactosidase level. e and f Testes in the experimental group received 0.3 μg of poly I:C, while those form the control group were injected with PBS. At various time points, testes were homogenized and examined for mRNA levels of TLR3 and RIG-I with qRT-PCR. g-j Sertoli and Leydig cells were separated from C57BL/6 mice and transfected with miR-26a mimic, miR-136 mimic, or NC mimic prior to treatment with poly I:C or PBS for 6 h. Total RNA was isolated from cells, and miR-26a, TLR3, RIG-I, and miR-136 levels were evaluated with qRT-PCR after normalization to GAPDH level. Results are expressed as means ± SEM (n = 3 replicates each experiment). *P < 0.05, **P < 0.01, ***P < 0.001