CREB-1 siRNA (h)

Protocol tips

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	Primary human umbilical vein endothelial cells (HUVECs), purchased from Lonza (Allendale, NJ), were maintained in endothelial basal medium (EBM) with EGM-2 Single Quots (Lonza, Allendale, NJ) as previously described. For cultured cells in defined normoxia (21% O2) or hypoxic conditions (1% O2), a modular incubator chamber (Forma Scientific) was used. CoCl2 was purchased from Sigma-Aldrich and added to the media (200 μM) for different periods of time (0, 6, 12 and 24 h). HUVECs were transfected with CREB siRNA (sc29281, Santa Cruz) or scrambled (Scr) siRNA (sc-37007, Santa Cruz) using Lipofectamine RNAi MAX Reagent (13778150, Thermo Fisher Scientific) according to the protocol.

Protocol tips

Primary human umbilical vein endothelial cells (HUVECs), purchased from Lonza (Allendale, NJ), were maintained in endothelial basal medium (EBM) with EGM-2 Single Quots (Lonza, Allendale, NJ) as previously described. For cultured cells in defined normoxia (21% O2) or hypoxic conditions (1% O2), a modular incubator chamber (Forma Scientific) was used. CoCl2 was purchased from Sigma-Aldrich and added to the media (200 μM) for different periods of time (0, 6, 12 and 24 h). HUVECs were transfected with CREB siRNA (sc29281, Santa Cruz) or scrambled (Scr) siRNA (sc-37007, Santa Cruz) using Lipofectamine RNAi MAX Reagent (13778150, Thermo Fisher Scientific) according to the protocol.

Publication protocol

Primary human umbilical vein endothelial cells (HUVECs), purchased from Lonza (Allendale, NJ), were maintained in endothelial basal medium (EBM) with EGM-2 Single Quots (Lonza, Allendale, NJ) as previously described. For cultured cells in defined normoxia (21% O2) or hypoxic conditions (1% O2), a modular incubator chamber (Forma Scientific) was used. CoCl2 was purchased from Sigma-Aldrich and added to the media (200 μM) for different periods of time (0, 6, 12 and 24 h). HUVECs were transfected with CREB siRNA (sc29281, Santa Cruz) or scrambled (Scr) siRNA (sc-37007, Santa Cruz) using Lipofectamine RNAi MAX Reagent (13778150, Thermo Fisher Scientific) according to the protocol.

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Manufacturer protocol

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