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2 protocols using ab137474

1

Piceatannol Inhibits Fibrosis Markers

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Piceatannol was purchased from Future Chem (Seoul, Korea). Anti-alpha smooth muscle actin (α-SMA; 1:1000, sc-130617), anti-CTGF (1:1000, sc-14939), anti-HDAC3 (1:1000, sc-11417), anti-HDAC4 (1:1000, sc-11418), anti-HDAC5 (1:1000, sc-133225), anti-TGF-β1 (1:1000, sc-146), anti-JNK (1:1000, sc-7345), anti-ERK1 (1:1000, sc-271269), and anti-GAPDH (1:1000, sc-32233) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Primary antibodies against collagen type I (1:1000, ab34710), HDAC2 (1:1000, ab12169), HDAC8 (1:1000, ab137474), and HDAC10 (1:1000, ab53096) were purchased from Abcam (Cambridge, MA, USA). Anti-fibronectin antibody (1:1000, MA5-11981) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Anti-HDAC1 antibody (1:1000, 06–720) was purchased from Merck Millipore (Darmstadt, Germany). Anti-HDAC6 (1:1000, 7612), anti-Smad3 (1:1000, 9523), anti-Smad2 (1:1000, 3103), anti-Smad4 (1:1000, 9515), anti-p-Smad3 (1:1000, 9520), anti-p-JNK (1:1000, 9251), anti-p-p38 (1:1000, 4511), anti-p-ERK1/2 (1:1000, 4370), and anti-p38 (1:1000, 8690) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
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2

Quantitative Analysis of Histone Deacetylases

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For western blot analysis, heart tissues were homogenized with a RIPA lysis buffer, as described previously.16) (link) Proteins were subjected to 8% or 10% SDS-PAGE and transferred to a PVDF membrane. The membranes were incubated in a 5% skim milk blocking solution for 1 h and then with the following primary antibodies: anti-HDAC1 (06-720, Millipore), anti-HDAC2 (ab12169, Abcam), anti-HDAC3 (sc-11417, Santa Cruz), anti-HDAC4 (sc-11418, Santa Cruz), anti-HDAC5 (sc-133225, Santa Cruz), anti-HDAC6 (#7612, Cell Signaling), anti-HDAC7 (3607-100, BioVision), anti-HDAC8 (ab137474, Abcam), anti-HDAC9 (3609-100, BioVision), anti-HDAC10 (ab53096, Abcam), and anti-GAPDH (sc-32233, Santa Cruz). The blots were incubated with anti-rabbit or anti-mouse horseradish-peroxidase-conjugated secondary antibodies for 1 h. The blots were developed using Immobilon™ Western Detection Reagents (Millipore, Billerica, MA, USA).
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