Fig. 3.

PGE₂ results in deacetylation of chromatin associated with the 15-PGDH gene promoter. (A) Chromatin immunoprecipitation (IP) of acetylated histone H3 (AcH3) is compared with IP of IgG (negative control) in cells treated with DMSO or PGE₂ (100 nM) for 24 h. Data represent average fold enrichment. *P < 0.05 compared with DMSO, Student’s t test. n = 3. (B) Representative immunoblot and quantitation of 15-PGDH levels in hCSCs treated with DMSO or HDACi SAHA (2.5 µM), TSA (1 µM), or HDAC-42 (1 µM) for 24 h. Acetylated histone H3(AcH3), positive control; β-actin, loading control. *P < 0.05 compared with DMSO, Student’s t test. n = 3. (C) Dose- (24 h) and time-dependent [SAHA (2.5 µM), TSA (1 µM), or HDAC-42 (1 µM)] increases in 15-PGDH after treatment with HDACi. *P < 0.05 compared with DMSO (C) or 0 time point (D), ANOVA, Dunn’s post hoc testing. n = 3. (E) Data represent fragments per kilobase of exon per million fragments mapped (FPKM) of class I, IIA, IIB, and IV HDACs expressed in hCSCs treated with DMSO or PGE₂ (100 nM) for 1 or 24 h mined from the RNA-seq dataset. GAPDH and RPLP0 are shown as controls (1/100th FPKM values). *P < 0.05 compared with corresponding DMSO control.