Fig 1: Protein levels of IL-6 (A, B), and MCP-1 (C, D) in BSMCs from asthma (A, C) and COPD (B, D) compared to BSMCs from healthy control groups were quantified by ELISA. n = 4 asthma, n = 3 healthy controls, n = 4 COPD, and n = 3 healthy control smokers. Data is representative of two independent experiments. One way ANOVA using Newman-Keuls multiple comparison test were performed to assess statistical significance between groups. Mean ± SE; (ns) p > 0.05, no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 2: Asprosin-induced inflammation in THP-1 macrophages is inhibited by TAK-242, a Toll-like receptor 4 (TLR4) inhibitor. (A) Tumour necrosis factor a (TNFa); (B) Interleukin-1ß (IL-1ß); and (C) IL-8 mRNA levels were measured by qRT PCR in THP-1 macrophages treated with 1 µM TAK-242 for 1 h, prior to 4 h stimulation with 100 nM asprosin or 100 ng/mL lipopolysaccharide (LPS). Data are presented as fold-change (mean ± SEM) in transcript. The experiments were repeated in three independent cultures. One-way ANOVA; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Secreted levels of TNFa (D), IL-1ß (E), MCP-1 (F) and IL-8 (G) were measured by a human cytokine BioPlex array in cell supernatant of THP-1 macrophages treated with 1µM TAK-242 for 1 h, prior to 4 h stimulation with 100 nM asprosin or 100 ng/mL LPS. The experiments were repeated in three independent cultures. One-way ANOVA; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.
Fig 3: Box-plot of each urinary biomarkers levels evaluated on ICU admission of COVID-19, according to death during hospital stay. (A) Urinary nephrin; (B) urinary MCP-1; (C) urinary KIM-1; (D) urinary NGAL.
Fig 4: MCP1/CCL2 present in conditioned media by TNFa-treated preadipocytes stimulates monocyte recruitment. THP-1 monocytes (1 × 105 cells) were subjected to a recruitment experiment following the same procedure that those in Figure 2. A group of these cells was subjected to migrate under CM stimulus (50%) from mammary preadipocytes treated or not treated with 20 ng/mL TNFa in the presence or absence of 10 µg/mL of anti-CCL2 blocking antibody (R&D system). (A) Representative photomicrographs (×20) of migratory cells in each condition and (B) bars represent mean ± SD of migrating cells per field. Friedman test, followed by Bonferroni’s multiple comparisons test, were used to derive all p values. * p < 0.05 and **** p < 0.0001. Data correspond to three independent experiments performed with CM from three different samples.
Fig 5: Soluble factors produced by TNFa-treated preadipocytes stimulate cancer epithelial migration in an MCP1/CCL2-dependent manner. (A,B) MCF-7 epithelial cells (2 × 105) were stimulated to migrate for a 48 h period against 50% of conditioned media with preadipocytes previously treated (72 h) or not treated with TNFa (20 ng/mL), using a Transwell system. CM CTL corresponds to a media conditioned in the absence of the factor. (C,D) MCF-7 epithelial cells were subjected to migrate in the same conditions as in A but included a group of cells that migrate in the presence of an anti-CCL2 blocking antibody (10 µg/mL). Bars represent mean ± SD of migrating cells per field. Friedman test, followed by Bonferroni’s multiple comparisons test, were used to derive all p values. * p < 0.05, *** p < 0.001 and **** p < 0.0001. Data correspond to three independent experiments performed with CM from three different samples.
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