Fig 1: Levels of NOS2 in the blood from patients with diabetes compared with healthy subjects. (A) Relative expression of NOS2 mRNA in the blood determined by reverse transcription-quantitative PCR. (B) Content of NOS2 protein in the blood determined by enzyme-linked immunosorbent assay. *P<0.05 vs. diabetes group. N=33. NOS2, nitric oxide synthase 2.
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-?, IL-1ß, iNOS, TNF-a, 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-? (B), IL-1ß (C), iNOS (D), TNF-a (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 ß-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.
Supplier Page from Abcam for Human iNOS ELISA Kit