Monacolin X Modulates VEGFR2 and PKC Signaling

Monacolin X was administered to HUVECs at IC₅₀ concentration and SU5416 (4 μM) for a duration of 12 h in the presence of TPA at a concentration of 10 nmol L⁻¹ for 4 h. The total RNA was isolated from HUVECs after 6 h of incubation with monacolin X. The amplification was performed using Clonetech SYBR Premix using the following set of primers: VEGFR2 forward: 5′-AGGAAGTAGCCGCATTTG-3′, reverse: 5′GGAGAAGACACAGACACA-3′; PKCα forward: 5′-TGGCAAAGGAGCAGAGAACT-3′, reverse: 5′-TGTAAGATGGGGTGCACAAA-3′; PKCη forward: 5′-AGTAGA CTGGTGGGCAATGG-3′, reverse: 5′-GATCCCTGTGG CATCTTCAT-3′; β-actin: forward: 5′-CTCTTCCAGCCTTCCTTCCT-3′, reverse: 5′-AGCACTGTGTTGGCGTACAG-3′. The PCR amplification was performed using qPCR (Applied Biosystem) under the following conditions: 40 cycles at 95 °C for 1 min, 61 °C for 1 min, and 74 °C for 2 min followed by 10 min at 74 °C. qPCR data were quantitatively analyzed by using the formation of 2-ΔΔCt. The relative expression levels of the mRNAs of the target genes were normalized using the β-actin internal standard.^{15 (link)}