Fig 2: Effect of miR-185 expression on the expression of NOS2. (A) Expression of miR-185 in HMEC-1 cells after transfection with agomiR-NC and agomiR-185. (B) Expression of NOS2 mRNA in HMEC-1 cells after transfection with agomiR-NC and agomiR-185. (C) Expression of NOS2 protein in HMEC-1 cells after transfection with agomiR-NC and agomiR-185. *P<0.05 and **P<0.01 vs. agomiR-NC group. N=3. miR, microRNA; NOS2, nitric oxide synthase 2; NC, negative control.

Fig 3: Plasma levels of CCAT1, IFN‐γ, IL‐1β, iNOS, TNF‐α, and IL10 in patients and controls. Plasma samples from 200 newly developed tuberculosis (N‐TB) patients, 102 recurrent tuberculosis (R‐TB) patients, and 102 controls were subjected to both quantitative reverse transcription polymerase chain reaction and enzyme‐linked immunosorbent assay to determine plasma levels of CCAT1 (A), IFN‐γ (B), IL‐1β (C), iNOS (D), TNF‐α (E), and IL‐10 (F). *p < 0.05

Fig 4: Macrophages control Mtb survival and promote osteogenic differentiation of MSCs via BMP/SMAD/RUNX2 pathway.a Lung CT scan from patients TB-7 and TB-3. Arrows indicate calcific densities in pathologic lesions. b, c Representative images of histologic analysis and Von Kossa staining (b), and p-SMAD1/5/8 and RUNX2 immunohistochemical analysis (c) of NC and TB lung sections from TB patients as in a. Single arrowheads, the osteoid with mass deposits of calcium (black); double arrowheads, dispersed deposits of calcium (brown); asterisks, the necrotizing foci. OC osteocyte; OB osteoblast; MGC multinucleated giant cell. Scale bars, 200 µm. d, e Quantitative analysis of p-SMAD1/5/8 (d) and RUNX2 (e) immunostaining as in c. f Immunoblot analysis of p-SMAD1/5/9, SMAD1/5/9, RUNX2, and β-Actin in six independent NC or calcified TB lung samples. g Immunoblot analysis of BMP2/4, p-SMAD1/5/9, SMAD1/5/9, RUNX2, and GAPDH in U937 cells. Cells were treated with 500 nM LDN-193189, 1 μM K02288, 10 μM Galunisertib or control DMSO, and infected with Mtb for 0–48 h. h Survival of Mtb in U937 cells treated as in g. i, j ELISA of TNF (i) and iNOS (j) from U937 cells treated with LDN-193189 or control DMSO and infected with Mtb as in g. k Representative images of CD29 and CD68 immunohistochemical analysis of NC and TB lung sections. Arrowheads indicate the gathering of CD29+ cells (indicating MSCs) and CD68+ cells (indicating macrophages). Scale bars, 200 µm. l Representative images of Alizarin Red S staining of BMSCs. BMSCs were cultured with conditional media derived from BMDMs (CM), Mtb-infected BMDMs (Mtb-CM), LDN-193189-pretreated and Mtb-infected BMDMs (LDN-Mtb-CM) or control media for 0–14 d. Arrowheads, the mineralized nodules. Scale bars, 200 µm. m– o Quantitative PCR analysis of Alp (m), Bglap (n), and Runx2 (o) mRNAs in BMSCs treated as in l. P > 0.05, not significant (ns); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (mean ± s.e.m. of n = 4 in d, e, h, and m–o, and n = 3 in i, j, two-way ANOVA). Results are representatives from at least three independent experiments.

Fig 5: Level of miR-185 in the blood. (A) Possible binding sites between miR-185 and NOS2 gene, as predicted by bioinformatics. (B) Relative expression of miR-185 in blood from patients with diabetes in contrast to control group. **P<0.01 vs. control group. N=33. miR, microRNA; NOS2, nitric oxide synthase 2.