Figure 4
5-aza increases the production of PGE2 from hMSCs through the up-regulation of synthesis enzymes.
(A-B) After treating hMSCs with 5-aza for 24 hr, COX2 and PTGES expression was detected through (A) real-time qPCR and (B) western blot analysis (C) After treating hMSCs with 5-aza for 24 hr, the changes in PGE2 expression levels were measured using ELISA. PGE2 secretion from hMSCs was increased by 5-aza treatment. (D) After treatment of 5-aza with or without the inhibition of COX2 (siCOX2), indirect-MLR was performed. The suppression of MLR by hMSCs was increased by treating with 5-aza and the effect was rebounded by inhibition of COX2. (E) Schematic diagrams indicating the locations of the promoter primers. (F) After treating hMSCs with 5-aza, bisulfide conversion and methyl-specific PCR were performed. In 5-aza treated hMSCs, the promoters of COX2, PTGES1 and PTGES2 were hypomethylated compared with control MSCs. pm, promoter locus; TSS, transcription start site; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Results show one representative experiment out of at least three. Results are shown as mean ± SD.
