Fig. 4
Pimecrolimus reduces PGE2 production by regulating COX2 expression in hUCB-MSCs. hUCB-MSCs were treated with a–d the indicated concentration or e 100 ng/ml of pimecrolimus for 72 h. a The levels of PGE2 in the cell culture supernatant of hUCB-MSCs were determined using an ELISA. B–c The expression of the COX2 mRNA was determined using b conventional PCR and c Quantitative Real-time PCR. d Western blotting data and quantification showed the relative expression levels of COX2 in hUCB-MSCs. Expression of COX2 was normalized against GAPDH housekeeping gene by ImageJ program. e The expression of COX2 in hUCB-MSCs was verified by immunostaining, Bar = 500 μm. In vitro experiments were performed in triplicate using hUCB-MSCs isolated from each donor (N = 3). The Kruskal–Wallis test with Dunn’s post hoc was used to compare the pimecrolimus-treated hUCB-MSCs with control group. n.s: not significant, *P < 0.05, **P < 0.01 and ***P < 0.001, ****P < 0.0001. The results are presented as the means ± SD