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. 2018 Sep 7;9:2037. doi: 10.3389/fimmu.2018.02037

Figure 10.

Figure 10

Direct targeting of pro-inflammatory cytokines by miR-125a-5p and let-7e-5p. Wild-type of MRE-mutated luciferase constructs based on the 3'UTR of TNFα (A), IL-6 (D), CCL7 (G), CCL3 (J), and CXCL8 (M) were cotransfected in HEK-293T cells with miR-125a-5p, let-7e-5p, miR-99b-5p mimics or with a negative control mimic. Results are expressed as mean (% variation ± SEM; n = 5) of the ratio between renilla luciferase and firefly control luciferase activities. Cell extracts from THP1 cells transduced with miR-125a OE, let-7e OE, and the control vector (CT; panels B,E,H,K,N) or with miR-125a-5p sponge, let-7e-5p sponge, and the control vector (CT; panels C,F,I,L,O) were stimulated for 6 h with LPS and then subjected to RIP assay using anti-Ago2 or IgG control Abs and levels of TNFα (B,C), IL-6 (E,F), CCL7 (H,I), CCL3 (K,L), and CXCL8 (N,O). Transcript levels were assayed in triplicate by Q-PCR (mean ± SEM; n = 4) and expressed as normalized fold enrichment in Ago2 IP (black columns) and leftover (white columns). (* < 0.05: ** < 0.01; *** < 0.001).