Nanog

Total RNA from MSC was extracted with TRIzol reagent (Life Technologies, CA, USA), and the RNA samples were converted into complementary DNA by using an Applied Biosystems High-Capacity cDNA Reverse transcription Kit (Invitrogen, MA, USA) according to the manufacturer's instructions. RT-PCR was performed using a QuantiTect SYBR Green PCR kit (Qiagen, Valencia, USA) and Corbett Research Rotor-Gene RG-3000 Real Time PCR System. Pre-designed primers for human GATA4, Nkx2.5, cTnI, Runx2, PPAR-γ, Col2a1, Nanog, Sox2, Oct4, Fog2, Art27, YAP, p21, and AREG were purchased from Bioneer (Daejeon, Korea). MiR-130a was quantified by RT-PCR using TaqMan® MicroRNA Reverse Transcription Kit (Life Technologies, USA). The primers of miR-130a and 18S were purchased from TaqMan® MicroRNA assay (Life Technologies, USA).

Synthetic small interfering RNAs (siRNAs) specific for GFP, EGFR, ATG7, NANOG, TRPV1, and CaMKKβ were purchased from Bioneer (Daejeon, KOR). The sequence were: non-specific GFP (green fluorescent protein), 5′-GCAUCAAGGUGAACUUCAA-3′ (sense), 5′-UUGAAGUUCACCUUGAUGC-3′ (antisense); EGFR #1, 5′-GAUCCACAGGAACUGGAUA-3′ (sense), 5′-UAUCCAGUUCCUGUGGAUC-3′ (antisense); EGFR #2, 5′-UUAGAUAAGACUGCUAAGGCAUAGG-3′ (sense), 5′-CCUAUGCCUUAGCAGUCUUAUCUAA-3′ (antisense); ATG7 #1, 5′-CAGCUAUUGGAACACUGUA-3′ (sense), 5′-UACAGUGUUCCAAUAGCUG-3′ (antisense); ATG7 #2, 5′-CAGCUAUUGGAACACUGUA-3′ (sense), 5′-UACAGUGUUCCAAUAGCUG-3′ (antisense); NANOG #1, 5′-GCAACCAGACCUGGAACAA-3′ (sense), 5′-UUGUUCCAGGUCUGGUUGC-3′ (antisense); NANOG #2, 5′-CUAAACUACUCCAUGAACA-3′ (sense), 5′-UGUUCAUGGAGUAGUUUAG-3′ (antisense); TRPV1 #1, 5′-GGAGACCUGUCUGCUGAAAUU-3′ (sense), 5′-AAUUUCAGCAGACAGGUCUUC-3′ (antisense); TRPV1 #2, 5′-GGAGUUCACCGAGAACUAU-3′ (sense), 5′-AUAGUUCUCGGUGAACUCC-3′ (antisense); CaMKKβ #1, 5′-GUGAAGACCAUGAUACGUA-3′ (sense), 5′-UACGUAUCAUGGUCUUCAC-3′ (antisense); and CaMKKβ #2, 5′-GACCAUCUGUACAUGGUGU-3′, and 5′-ACACCAUGUACAGAUGGUC-3′ (antisense). For in vitro delivery, the cells were transfected with 100 pmol of synthesized siRNAs using Lipofectamine 2000 (Invitrogen, San Jose, CA, USA, 11668027) according to the manufacturer’s instructions.