RNA isolation and quantitative real-time PCR (qPCR) were performed as described in previous studies [
20 (link),
21 (link) ]. Briefly, the RNA samples were collected from BMDMs using TriZol (Invitrogen) according to the manufacturer’s instructions. Approximately 2 μg of total RNA was reverse transcribed into cDNA using a Reverse Transcription kit (Bio-Rad Laboratories, Hercules, USA). Individual real-time PCR was performed using the SYBR Green PCR Master Mix (Bio-Rad Laboratories) on a MyiQ Single Colour Real-time PCR Detection System (Bio-Rad Laboratories). The primer sequences used forqPCR are listed in
Supplementary Table S2 . The relative messenger RNA (mRNA) levels were calculated using the 2
-ΔΔCT method and
β-actin was used as an internal reference.
20 (link),
21 (link) ]. Briefly, the RNA samples were collected from BMDMs using TriZol (Invitrogen) according to the manufacturer’s instructions. Approximately 2 μg of total RNA was reverse transcribed into cDNA using a Reverse Transcription kit (Bio-Rad Laboratories, Hercules, USA). Individual real-time PCR was performed using the SYBR Green PCR Master Mix (Bio-Rad Laboratories) on a MyiQ Single Colour Real-time PCR Detection System (Bio-Rad Laboratories). The primer sequences used forqPCR are listed in
-ΔΔCT method and
β-actin was used as an internal reference.
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