Abi 7900 ht fast real time cycler

Total RNA was extracted from the indicated cells or mouse colon tissues with TRI reagent (Sigma) according to the manufacturer’s instructions. RNA samples were reverse-transcribed into cDNA with a GoScript^TM Reverse Transcription kit (Promega). The cDNA samples were amplified by real-time PCR with a SYBR Green kit (Toyobo) on an ABI 7900 HT Fast Real-Time cycler (Applied Biosystems). The expression of target genes was normalized to expression of housekeeping gene beta actin. In Figs. 2h and 6h, arbitrary units (AU) were introduced to normalize the difference between different batches of samples. Primers used were listed in Supplementary Table 3.

Intestinal tissue DNA was extracted with TIANamp Micro DNA Kit (TIANGEN, Cat. No. DP316, Beijing, China). qRT-PCR was performed on DNA with Premix Ex Taq^TM (Probe qPCR) (Takara, Cat. No. RR390A, Kusatsu, Japan) in a 20-μL volume on an ABI 7900 HT Fast Real-Time cycler (Applied Biosystems, Foster city, USA) according to the manufacturer’s instruction using the housekeeping gene GAPDH (for mouse) as a control. The specific primers of GAPDH and P. distasonis are from previous studies [69 (link), 70 (link)] and listed in Table S3.

Total RNA was extracted from homogenized lung or BMDMs with TRIzol reagent (SIGMA) according to the manufacturer’s instructions. Synthesis of cDNA was performed with a GoScript^TM Reverse Transcription kit (Promega). Real time quantitative PCR was performed with the SYBR Green qPCR Master Mix (TOYOBO) on an ABI 7900 HT Fast Real-Time cycler (Applied Biosystems). The expression of target genes was normalized to expression of housekeeping gene Gapdh (glyceraldehyde-3-phosphate dehydrogenase). The qPCR primers used in this study were listed in Supplementary Table 1.