We extracted total RNA from BMDMs using QIAzol Lysis Reagent (miRNeasy Micro Kit; QIAGEN). The first strand of cDNA was synthesized from 100ng RNA with SuperScript VILO cDNA synthesis kit (Invitrogen), and then RT-qPCR performed on an ABI 7500 Fast real-Time PCR System (Applied Biosystem) using TaqMan universal PCR Master Mix II with UNG. The TaqMan Gene Expression Assay Mix for mouse Cd36 (Mm00432403_m1), Axl (Mm00437221_m1), Tnf (Mm00443258_m1) and Gapdh (Mm99999915_g1) were obtained from Thermo Fisher Scientific. The cycle time values of candidate gene were normalized to endogenous control Gapdh in the same sample. The expression level of mRNA was calculated using delta delta CT (∆∆CT) method as previously described.3 (link), 10 (link), 17 (link)
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Quantifying gene expression in BMDMs
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Other organizations : National Taiwan University, Yale University, Universidade de São Paulo
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Variable analysis
independent variables
- Not explicitly mentioned
- MRNA expression levels of Cd36, Axl, Tnf
- Gapdh (used as an endogenous control)
- No positive or negative controls were explicitly mentioned in the provided information.
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