Through our GO network analysis, 17 of 73 genes, i.e., ATP binding cassette subfamily A member 13 (ABCA13), actin alpha2 (ACTN2
), ATPase phospholipid transporting 9A (ATP9A), C2 calcium dependent domain containing 3 (C2CD3), centrosomal protein 152 (CEP152), growth hormone (GH), glucagon like peptide 2 receptor (GLP2R), GNAS complex locus (GNAS), interleukin 22 receptor subunit alpha 2 (IL22RA2), microtubule associated protein 1 light chain 3 beta (MAP1LC3B), I-BAR domain containing (MTSS1), olfactomedin 1 (OLFM1), slowmo homolog 2 (SLMO2), prominin 1 (PROM1), sperm associated antigen 5 (SPAG5), transforming growth factor beta receptor 3 (TGFBR3), ubiquitin specific peptidase 36 (USP36), were found associated with six major GO terms.
The results showed that the pluripotency gene POU5F1 (POU class 5 homeobox 1) decreased, while the typical cardiac genes, TNNT2 (troponin T type 2), MYH6 (myosin heavy chain 6), MYL2 (myosin regulatory light chain 2), NKX2.5 (NK2 homeobox 5), and ACTN2 (actinin alpha 2), increased over time (Figure S1).
The qPCR analysis was carried out to assess the relative mRNA expression (normalized with GAPDH) of the pluripotency gene POU5F (A) and the cardiac genes MLY2 (B), TNNT2 (C), NKX2.5 (D), ACTN2 (E), and MYH6 (F) at different times after the onset of differentiation.
Using the STRING software, COX5B and MYH6 were found to belong to two separated networks but linked with a level of confidence of 0.508 through cytochrome c oxidase subunit 6 A2 (COX6A2) and actinin alpha 2 (Actn2
) (Figure 3, Tables 3 and 4).
As for SHED cardiac induction, only a handful of genes, namely, GATA4, HAND2, ADRB1, NPPA, PLN, SLC8A1, ACTN2
, MLY2, TNNT2, MB, and CKM, were upregulated significantly (P < 0.05) at the 7th day.
Genes for sports doping and physiologic response Gene/Product System/ target Gene product Physiologic tissue properties response ACE Skeletal Peptidyl ACE-I seems to muscles dipeptidase correlate with enduranceACE-D in?volved in sprint and powers ACTN2
,3 Muscular system Actin-binding Involved in fast prote?ins related twitch muscles to dystrophin Endorphins, Central and Widely active Pain modulation.
Next, D13 EBs were co-stained with STAT3 and cardiac specific marker Actn2
to examine the expression of STAT3 when Dox was added on D0 of cardiomyocyte differentiation of ESCs.