Figure 3.

The Effect of miR-139 and miR-192 on Macrophage Polarization In Vitro

(A) BMDMs were transfected with 100 nM scramble control or inhibitors against miR-139 or miR-192 for 48 h, followed by stimulation with LPS (100 ng/mL) plus IFN-γ (20 ng/mL) or IL-4 (20 ng/mL) for an additional 48 h. The mRNA levels of iNOS and Arg1 were determined by qRT-PCR. (B–E) BMDMs were transfected with 100 nM control or miR-192 mimic for 48 h, followed by stimulation with LPS (100 ng/mL) plus IFN-γ (20 ng/mL) or IL-4 (20 ng/mL) for an additional 48 h. (B and C) Supernatants from the two groups of cells were analyzed for TNF-α (B) and IL-12 (C) by ELISA. (D and E) RNA was extracted from the two groups of cells and subjected to qRT-PCR analysis with primers specific to iNOS (D) for M1 polarization or Arg1, YM1, and FIZZ1 (E) for M2 polarization. GAPDH was set as the endogenous control. The data are expressed as the mean ± SEM of three independent experiments. NC, negative control. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.