18s ribosomal rna 18s

mRNAs were extracted from ventricular tissues or cell lysates using RNeasy Mini kits (Qiagen, Cat. No. 74104, Hilden, Germany). Reverse transcription was performed using an iScript cDNA synthesis kit (Bio Rad, Cat. No. 1708891, Hercules, CA, USA). PCR reactions were performed with an ABI Step One Real-Time PCR System (Applied Biosystems, Quant Studio3, Foster, CA, USA) using Fast SYBR Green Real-time PCR Mixture (Applied Biosystems, Cat. No. 4385612, Foster, CA, USA) [52 (link)]. Primers for CTGF (Takara Bio Inc. Cat. No. MA028643), TGF-β (Takara Bio Inc. Cat. No. MA122075), collagen I (Takara Bio Inc. Cat. No. MA128559), collagen III (Takara Bio Inc. Cat. No. MA126692), OPN (Takara Bio Inc. Cat. No. MA114392) (Takara Bio Inc. Cat. No. HA201672), MCP-1 (Takara Bio Inc. Cat. No. MA066003), HGF (Takara Bio Inc. Cat. No. MA105799), and 18S ribosomal RNA (18S) (Takara Bio Inc. Cat. No. MA050364) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Takara Bio Inc. Cat. No. HA067812) are available commercially. The mouse heart samples were normalized to values for 18s mRNA expression (ΔΔCT method). The human cardiomyocyte culture samples were normalized to values for GAPDH mRNA expression (ΔΔCT method).

mRNAs were extracted from macrophages using RNeasy Mini Kits (QIAGEN, Cat. No. 74104). Reverse transcription was performed using an iScript cDNA Synthesis Kit (Bio-Rad, Cat. No. 1708891). qPCR was performed with an ABI Step One Real-Time PCR System (Applied Biosystems, QuantStudio 3) using a Fast SYBR Green Real-time PCR Mixture (Applied Biosystems, Cat. No. 4385612) [27 (link)]. Primers for monocyte chemotactic protein 1 (MCP-1) (Takara Bio Inc., Cat. No. MA066003), interleukin 1 beta (IL-1β) (Takara Bio Inc., Cat. No. MA025939), inducible nitric oxide synthase (iNOS) (Takara Bio Inc., Cat. No. MA083369), matrix metalloproteinase- (MMP-) 2 (Takara Bio Inc., Cat. No. MA127110), MMP-9 (Takara Bio Inc., Cat. No. MA031311), and 18S ribosomal RNA (18S) (Takara Bio Inc., Cat. No. MA050364) were commercially available. Each sample was normalized to values for 18S mRNA expression (ΔΔCT method) [27 (link)].