Fig. 3: JMJD3 enhances TGF-β1-induced Smad2 and Smad3 activation via induction of syntenin.
From: Epigenetic regulation of TGF-β-induced EMT by JMJD3/KDM6B histone H3K27 demethylase
a Phosphorylation of endogenously expressed smad2 and smad3 in BZR cells transfected with control shRNA or JMJD3 shRNA in response to TGF-β1 (5 ng/ml) for the indicated periods of time. b Expression level of p-smad2 and p-smad3 in A549 cells transfected with JMJD3 siRNA in response to TGF-β1 (5 ng/ml) for 1 h. c Expression level of p-smad2 and p-smad3 in BEAS-2B cells transfected with HA-tagged JMJD3 expression vector in response to TGF-β1 (5 ng/ml) for 1 h. d Luciferase activity of SBE-Luc reporter plasmid-expressed BZR cells transfected with control siRNA or JMJD3 siRNA in response to TGF-β1 (5 ng/ml) for 24 h. Columns, mean of three independent experiments performed in triplicate; bar, S.D.; ***P < 0.001 versus control, n.s.: nonsignificant. e Chromatin immunoprecipitation (ChIP) assay to analyze smad3-binding to target promoters such as CTGF (left panel), snail (middle panel), and slug (right panel) in BZR cells transfected with control siRNA or JMJD3 siRNA in response to TGF-β1 (5 ng/ml) for 1 h. Columns, mean of two independent experiments performed in triplicate; bar, S.D.; ***P < 0.001 versus control, n.s.: nonsignificant. f Real-time RT-PCR of CTGF (left panel) and smad7 (right panel) expression levels in A549 cells transfected with control siRNA or JMJD3 siRNA in response to TGF-β1 (5 ng/ml) for 12 h. Columns, mean of three independent experiments performed in triplicate; bar, S.D.; *P < 0.05, ***P < 0.001 versus control, n.s.: nonsignificant.
