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. Author manuscript; available in PMC: 2017 Feb 8.
Published in final edited form as: Nat Microbiol. 2016 Aug 8;1(10):16132. doi: 10.1038/nmicrobiol.2016.132

Figure 2. CAF inhibits LPS-induced miR-301b expression by regulating cAMP/PKA/CREM/NF-κB axis.

Figure 2

(a) LPS (100 ng/ml) induces miR-301b expression in MH-S, MLE-12, and BMDM cells in a time dependent manner as determined by qRT-PCR. (b) Northern blot showing the expression of miR-301b, 15a, and 301a in cells treated with LPS (100 ng/ml, 6 h). (c) qRT-PCR of LPS-induced miR-301b expression in BMDM from TLR4−/− and TLR-2−/− mice, or in MH-S and MLE-12 cells with TLR4, MyD88 or TRIF knockdown. (d) Cells were cultured for 6 h with LPS in combination with FR180204 (0.3 μM), SB203580 (0.5 μM), SP600125 (90 nM) and SN50 (10 μM). miR-301b was analyzed by qRT-PCR. (e) After CAF pretreatment for 24 h, LPS (100 ng/ml) was added for another 6 h incubation and miR-301b was detected by qRT-PCR. (f) Cells were treated with LPS in combination with Sp-8-Br (2.5 μM) and Rp-8-Br (4 μM) for 6 h and miR-301b was analyzed by qRT-PCR. Panel a, c–f, means+SD from triplicate; b, representative of 3 independent experiments. *, p<0.05, **, p<0.01. One-way ANOVA with Tukey’s post hoc.