RNA preparation was carried out in cells by direct lysis using the Trizol–Chloroform method, following the manufacturer’s protocol.
Reverse transcription was carried out using the MMLTV reverse transcriptase kit from Invitrogen (Thermo-Fisher, Bd Sébastien Brant, Parc d’Innovation, France).
The expression of the CD24 gene was quantified by TaqMan RT-PCR utilizing the Applied Biosystem StepOne Plus cycler (Applied Biosystems, Austin, TX, USA) and TaqMan Gene Expression Assay with primer and probe sets (Applied Biosystems) for CD24 (Hs02379687_s1).
HPRT and WYHAZ (ABI, Branchburg, NJ, USA) were used to standardize the expression level. The relative amount of CD24 was calculated by employing the comparative CT method (2−ΔΔCT) [49 (link)]. Amplification was performed for the JEG-3 and BeWo cells transfected by STOX1-A or STOX1-B polymorphic variants compared with the empty expression vector used as a mock control.
Reverse transcription was carried out using the MMLTV reverse transcriptase kit from Invitrogen (Thermo-Fisher, Bd Sébastien Brant, Parc d’Innovation, France).
The expression of the CD24 gene was quantified by TaqMan RT-PCR utilizing the Applied Biosystem StepOne Plus cycler (Applied Biosystems, Austin, TX, USA) and TaqMan Gene Expression Assay with primer and probe sets (Applied Biosystems) for CD24 (Hs02379687_s1).
HPRT and WYHAZ (ABI, Branchburg, NJ, USA) were used to standardize the expression level. The relative amount of CD24 was calculated by employing the comparative CT method (2−ΔΔCT) [49 (link)]. Amplification was performed for the JEG-3 and BeWo cells transfected by STOX1-A or STOX1-B polymorphic variants compared with the empty expression vector used as a mock control.
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