Data on the gene expression of cardiomyocyte exposed to hypothermia

Hypothermia is widely used in neurosurgery and cardiac surgeries. However, little is known about the underlying molecular mechanisms. We previously reported that the transcriptome responses of cardiomyocyte exposed to hypothermia, “The transcriptome responses of cardiomyocyte exposed to hypothermia” [4]. Herein, we provide the hypothermia inhibited proliferation of cardiomyocyte cells in vitro and the details of transcription factors in regulation of differentially expressed genes.


a b s t r a c t
Hypothermia is widely used in neurosurgery and cardiac surgeries. However, little is known about the underlying molecular mechanisms. We previously reported that the transcriptome responses of cardiomyocyte exposed to hypothermia, "The transcriptome responses of cardiomyocyte exposed to hypothermia" [4]. Herein, we provide the hypothermia inhibited proliferation of cardiomyocyte cells in vitro and the details of transcription factors in regulation of differentially expressed genes. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Value of the data
The data provides the inhibition of hypothermia on the cardiomyocytes in vitro culture. This data provides the details of differentially expressed genes (DEGs) of cardiomyocytes exposed on hypothermia.
The data may stimulate further research on the function of transcription factor (TF) stimulated in cardiomyocytes under hypothermia.

Data
The viable cell number was determined by Cell Counting Kit-8 (CCK-8) assay ( Fig. 1). As shown in Fig. 1, the relative cell number of hypothermia was decreased as the time of hypothermia culture.
The details of changed genes are listed in Supplementary Table S1. Pathway enrichment analysis was performed considering the notion that different genes cooperate with each other to exercise their biological functions. The changed pathways are listed in Supplementary Table S2.
The details of TFs in regulation of DEGs are listed in Supplementary Table S3. Primers for the 11 randomly selected differentially expressed genes are listed in Supplementary  Table S4. The amount of 18s, a constitutive transcript (endogenous control) was normalized to check the fold change in the expression of the target genes (Fig. 2).

Experimental design and hypothermia treatment
AC16 human adult ventricular cardiomyocytes were cultured in incubator with normal temperature (37°C) and 5% CO 2 . The hypothermia treated AC16 cells were cultured in another incubator with low temperature (28°C) and 5% CO 2 . The cells incubated for six hours and the RNA was extracted using the TRIzol (Invitrogen) reagent. This experiment was repeated three times (N ¼3). Then, the RNA was isolated by Genminix Co. (Shanghai, China). Finally, the microarray hybridization was completed [4].

Cells culture
AC16 human adult ventricular cardiomyocytes were purchased from the American Type Cell Culture (ATCC) [2]. The cells were maintained in DMEM/F-12 supplemented with 10% fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. Cells were cultured at 37°C or 28°C with 5% CO 2 .

Cell Counting Kit-8 (CCK8) assay
Cell viability was determined by the Cell Counting Kit-8 assay (DOJINDO, Japan), according to the manufacturer's instructions. The absorbance of each well was measured at 450 nm with a microtiter plate reader.

Microarray hybridization for the hypothermia regulated transcriptome
Microarray hybridization has been described previously in [4].

GO analysis
Gene Ontology (GO) analysis was applied to analyze the main function of the differentially expressed genes according to the gene ontology, which is the key functional classification of NCBI that can organize genes into hierarchical categories and uncover the gene regulatory network based on biological process and molecular function [1].

Pathway analysis
Pathway analysis was used to identify the significant pathway of the differential genes according to KEGG, Biocarta and Reactome. Fisher's exact test was used to select the significant pathway, and the threshold of significance was defined by P-value and FDR. The enrichment Re was calculated based on the previously published equation [3].

Statistical analysis
RVM t-test was applied to filter the differentially expressed genes for the control and hypothermia treated group because the RVM t-test can raise degrees of freedom effectively in the cases of small