Fig 1: Enzyme-linked immunosorbent assay results of candidate proteins in MII and Gv oocytes in follicular fluid. (A) Prostatic acid phosphatase (ACPP); (B) CD5 antigen-like (CD5L); (C) Alpha 2 macroglobulin (A2M); (D) Peptidyl-prolyl cis-trans isomerase A (PPIA); (E) Beta-actin (ACTB). *** P < 0.0001.
Fig 2: Serum Cyclophilin A (CypA) levels in control subjects (n = 44) and in patients (n = 40) with coronary artery disease (CAD). (A) Frequency plot: Data represented as CypA levels (X-axis) versus number of subjects (n = 84) (Y-axis). (B) Data are shown as dots of CypA levels in control subjects (control, n = 44) and CAD patients (n = 40). (C) Data are shown as box-and-whisker plots of CypA levels in control subjects (n = 44) and CAD patients (n = 40). CypA was significantly elevated (***P < 0.001, Mann-Whitney U test) in CAD patients compared with controls.
Fig 3: Effect of compounds 1, 2, and GraL over intra and extracellular CypA, B and C levels in human T lymphocytes stimulated with Con A Cells were pre-treated with compound 1, 2 (0.1 µM), GraL (1 μM) or CsA (0.2 µM) for 2 h and then were stimulated with Con A (50 μg/mL) for 48 h. CypA (A), CypB (B), and CypC (C) levels were measured by ELISA in the medium of T lymphocytes. The line represents the unstimulated cells values. The intracellular expression of CypA (D), CypB (E), and CypC (F) was measured by western blot in cytosolic lysates of T lymphocytes. Band intensity was normalized by β-actin. Data are the result of average ± SEM (n = 3). The values are shown as the difference between cells treated with Con A and cells treated with compounds plus Con A, *p < 0.05, **p < 0.01 and ***p < 0.001. Values of cells treated with Con A were compared with control cells, # p < 0.05, ## p < 0.01 and ### p < 0.001. One-way ANOVA test with Dunnet’s post hoc analysis and Student´s t-test were used for statistical analysis.
Fig 4: Intra and extracellular CypA, B, and C levels in human T lymphocytes stimulated with Con A Cells were treated with Con A (50 μg/mL) for 48 h. CypA (A), CypB (B), and CypC (C) levels were measured by ELISA in the medium of T lymphocytes. The intracellular expression of CypA (D), CypB (E), and CypC (F) was measured by western blot in cytosolic lysates of T lymphocytes. Band intensity was normalized by β-actin. Data are the result of average ± SEM (n = 3). The values are shown as the difference between cells treated with Con A and untreated cells, *p < 0.05, *p < 0.01 and ***p < 0.001. One-way ANOVA test with Dunnet’s post hoc analysis and Student´s t-test were used for statistical analysis.
Fig 5: Effect of compounds 1, 2, and GraL over human T lymphocytes migration. Cells were pre-treated with compound 1, 2 (0.1 µM), GraL (1 μM) or CsA (0.2 µM) for 2 h and then were stimulated with Con A (50 μg/mL) for 48 h. Then, T cells were placed in the migration chamber for 24 hours in the presence of the chemoattractant CypA (50 ng/mL) (A) or CypB (5 ng/mL) (B). Controls of untreated T cells in the presence or absence of Cyps are also included. Data are the result of average ± SEM (n = 3). The values are shown as the difference between Con A migratory cell numbers compared to cells treated with compounds plus Con A, *p < 0.05 and **p < 0.01. Values of cells treated with Con A were compared with cells in the presence of CypA or B, # p < 0.05. Values of untreated cells in absence of CypA or B were compared with untreated cells in presence of CypA and B, + p < 0.05 or ++ p < 0.01. One-way ANOVA test with Dunnet’s post hoc analysis and Student´s t-test were used for statistical analysis.
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