Lipofectamine® 2000 Transfection Reagent

Protocol tips

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	Final concentration of miRNA mimic and siRNA was 30 nM. change media from antibiotics media to decidual differentiation with 0.5 mM 8-br-cAMP and 10−6 M MPA or control media.

Protocol tips
Final concentration of miRNA mimic and siRNA was 30 nM. change media from antibiotics media to decidual differentiation with 0.5 mM 8-br-cAMP and 10−6 M MPA or control media.

Publication protocol

mmediately prior to transfection, the culture medium of HESCs was changed to antibiotic-free decidualization medium containing 0.5 mM 8-br-cAMP and 10−6 M MPA. Control cultures were maintained at the standard medium. mirVanaTM miRNA mimic (#MC11340; Applied Biosystems) for hsa-miR-542-3p or siRNA targeting IGFBP1 (Silencer select, s7225; Applied Biosystems) was mixed with Lipofectamine 2000 (Invitrogen), according to the manufacturer’s instructions, and added to HESCs in six-well culture plates at a density of 80–90% confluence. Control cultures were transfected with control miRNA mimic or non-targeting siRNA. Final concentration of miRNA mimic and siRNA was 30 nM. After 6 h and 3 days, cells were subjected to a change of antibiotics media to decidual differentiation with 0.5 mM 8-br-cAMP and 10−6 M MPA or control media. Morphological evaluation, RNA extraction and qRT-PCR analysis were performed on day 6 of decidualization.

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Manufacturer protocol

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