Fig 2: 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 3: 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.

Fig 4: 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 5: Concentration of cyclophilins B and D throughout the menstrual cycle. (a) Box-and-whisker plot of CypB concentrations in each phase. (b) Box-and-whisker plot of CypD concentrations in follicular, periovulatory and mid-luteal phases. Cyps concentrations were determined by ELISA kits (n = 11). Statistical differences were analysed with Friedman and Dunn-Bonferroni tests, **p < 0.01.