Silencing HDAC2 and HDAC3 to Regulate PPARγ

HDAC2 and HDAC3 siRNA duplex (Guangzhou RiboBio Company Limited) were respectively used to interfere with endogenous HDAC2 and HDAC3 mRNA expression. Transfection of siRNA was carried out as we have described in detail previously (Zhang et al., 2017 (link)). The following siRNA oligos were used: HDAC2: 5′-TCCGTAATGTTGCTCGATG-3′; HDAC3: 5′-GCATTGATGACCAGAGTTA-3′. The non-specific siRNA (scrambled siRNA; Guangzhou RiboBio Company Limited) was used as control. The expression of PPARγ mRNA and protein were measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and Western blot analyses, respectively. The levels of Ace-H3K9 in the promoter region of LPL, PPARγ, and miR-29a were investigated by Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay.

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Zhang J., Liu Y., Wang S., Que R., Zhao W, & An L. (2020). Exploration of the Molecular Mechanism for Lipoprotein Lipase Expression Variations in SH-SY5Y Cells Exposed to Different Doses of Amyloid-Beta Protein. Frontiers in Aging Neuroscience, 12, 132.

Publication 2020

HDAC2 HDAC3 siRNA duplex scrambled siRNA

Chip assay Mrna Oligos Pparγ Protein Reverse transcriptase polymerase chain reaction Sirna Transfection Western blot analyses

Corresponding Organization : China Medical University

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Variable analysis

independent variables

HDAC2 siRNA duplex
HDAC3 siRNA duplex

dependent variables

Expression of PPARγ mRNA
Expression of PPARγ protein
Levels of Ace-H3K9 in the promoter region of LPL, PPARγ, and miR-29a

control variables

Non-specific siRNA (scrambled siRNA)

controls

Positive control: Not specified
Negative control: Non-specific siRNA (scrambled siRNA)

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