Spinsmart rna mini purification kit

Total RNA was isolated by using SpinSmart RNA Mini Purification Kit (Denville Scientific Inc., Metuchen, NJ, https://www.denvillescientific.com). One microgram of RNA was reverse transcribed by using a High Capacity RNA‐cDNA kit (Thermo Fisher). Real‐time quantitative PCR was performed by using Perfecta SYBR Green FastMix on a Stratagene MX3000P (Thermo Fisher).
The following primers were used: COL1‐FOR primer 5′CTG CTG GCA AAG ATG GAG A3′ and COL1‐REV primer 5′ACC AGG AAG ACC CTG GAA TC3′; COL3‐FOR primer 5′ CAA ATG GCA TCC CAG GAG3′ and COL3‐REV primer 5′CAT CTC GGC CAG GTT CTC 3′; α‐SMA‐FOR primer 5′AGC GTG AGA TTG TCC GTG ACA T3′ and α‐SMA‐REV primer 5′ GCG TTC GTT TCC AAT GGT GA3′; VE‐cadherin‐FOR primer 5′ACC ATC GCC AAA AGA GAG AC3′ and VE‐cadherin‐REV primer 5′TCT TGC CAG CAA ACT CTC CT3′; CD31‐FOR primer 5′AGC GCA GTC TTA CCG AAG G3′; CD31‐REV primer 5′TCT TGC CAG CAA ACT CTC CT3′; Oct4–FOR primer CAAGGCAAGGGAGGTAGACA; REV primer GCTCCTGATCAACAGCATCA; KLF4 – FOR primer CCAGCAAGTCAGCTTGTGAA; REV primer GGGCATGTTCAAGTTGGATT; c‐Myc – FOR primer GCCTAACCTCACAACCTTGG; REV primer CCTATTTACATGGGAAAATTGGA; Nanog – FOR primer AGCCTCCAGCAGATGCAAGA; and REV primer GCACTTCATCCTTTGGTTTTGA. Values were normalized for the abundance of the amplified 18s rRNA in each experiment. Fold change in gene expression was determined by using the 2‐ΔΔCT method.