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. 2021 Oct 14;12:756825. doi: 10.3389/fimmu.2021.756825

Figure 2.

Figure 2

miR-4512 directly targets TLR4 and CXCL2. (A, B) Luciferase activity of WT/MUT-CXCL2 and WT/MUT-TLR4 plasmids in HEK293T following transfection with an miR-4512 agomir. Data are presented as the mean ± SEM. ****P < 0.0001; ***P < 0.001; NS = not significant. (C–F) RT-qPCR analysis of CXCL2 and TLR4 transcript levels in human primary monocytes and macrophages 48 h after transfection with NC/miR-4512 agomiR/antagomiR. Data are presented as the mean ± SEM. ****P < 0.0001; ***P < 0.001. (G, H) CXCL2 concentration in the supernatants obtained from human primary monocytes (G) and macrophages (H) 48 h after transfection with NC/miR-4512-agomir/antagomir. Data are presented as the mean ± SEM. ***P < 0.001; **P < 0.01; *P < 0.05; NS, not significant. (I) Western blot analysis of TLR4 expression in monocytes and macrophages 72 h after transfection with NC/miR-4512-agomir/antagomir. Results are representative of three independent experiments, with actin used as an internal control. (J, K) CXCL2 concentration in the plasma of SLE and control subjects from Tibetan (J) and Han (K) populations. Data are presented as the mean ± SEM. ****P < 0.0001; ***P < 0.001. (L) Western blot analysis of TLR4 expression in PBMCs isolated from SLE patients (A1–A4) and healthy controls (B1–B4). Results are representative of three independent experiments.