Figure 3.

ConA triggers MIF expression and secretion in human hepatoma cells. HuH-7 and Hep G2 cells were treated with or without ConA (20 μg/ml) for indicated time periods. (a) The MIF mRNA level was determined by RT-PCR. (b) The level of MIF secretion from culture supernatant was detected by MIF-ELISA (left y axis). △MIF represents MIF concentration in ConA-treated media compared with the control group. The cytotoxicity was analyzed by LDH release activity from culture supernatant (right y axis). LDH release (% of ctrl) represents the percentage of LDH activity in media compared with the control group. (c) HuH-7 cells were treated with or without ConA (20 μg/ml) for indicated time points in the presence or absence of cycloheximide (CHX) (10 μM) pre-treatment for 30 min. The level of MIF from culture supernatant was detected by MIF-ELISA. All data are presented as the mean±S.D. from at least triplicate independent experiments; *P<0.05, **P<0.01, ***P<0.001, ns indicates no significance