Immunosuppressive Factor MNSFβ Regulates Cytokine Secretion by Mouse Lymphocytes and Is Involved in Interactions between the Mouse Embryo and Endometrial Cells In Vitro
Figure 5
Action of culture supernatants from the HCC-94 cells, treated with MNSFβ overexpression, siRNA, or immunoneutralizing action of anti-MNSFβ, on the secretion of IL-4 and TNFα from mouse lymphocytes. (a, c, e) IL-4 concentrations in mouse lymphocytes culture supernatants detected by ELISA. (b, d, f) TNFα concentrations in mouse lymphocyte culture supernatants detected by ELISA. The time-point when HCC-94 cells culture supernatants were added into the mouse lymphocytes culture was designated as 0, and lymphocyte culture supernatants were sampled at 6, 12, and 24 hours, respectively. MNSFβ: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDsRed-N1/MNSFβ; NC for MNSFβ: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDsRed-N1; HCC-94: lymphocytes cultured in supernatant of HCC-94 cells without plasmid transfection (as negative control); siRNA: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDC316-EGFP-U6/MNSFβ-P1; “NC for siRNA”: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDC316-EGFP-U6/MNSFβ-NC. MNSFβ + antibody: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDsRed-N1/MNSFβ that were beforehand incubated with anti-MNSFβ antibody; “MNSFβ + preimmune”: lymphocytes cultured in supernatant of HCC-94 cells transfected with pDsRed-N1/MNSFβ that were previously incubated with rabbit preimmune sera (control); Antibody: only rabbit anti-MNSFβ sera added to the mouse lymphocytes culture (control). Asterisks (*) indicate the significance (P < 0.05) in comparison with “NC for MNSFβ” (a, b), “NC for siRNA” (c, d), and “MNSFβ + preimmune” (e, f).
