Gene expression profile at single cell level of periheral blood mono-nucleus cells (PBMC) in response to bacterial protein (PFOR) stimulation

We identified Pyruvate:Ferredoxin Oxidoreductase (PFOR) as a bioactive protein from Faecalibacterium prausnitzii. We usd single cell transcriptome sequencing to fully evaluate the PFOR's effect on PBMC. Overall design: In this study, PBMCs samples were treated with the PFOR protein or its 2nd inactive peptide (as control) respectively. The scRNA-seq of 10× Genomics based on droplet capture technology was done by Novogene (Tianjin, China) according to their commercially standard protocol (Ni et al., 2021). Briefly, the harvested single cells and the gel beads with barcoded sequences and oil droplets were put in the machine together. These gel beads precisely capture the target cells, then cells were lysed, thus releasing a large quantity of RNA and barcoded sequences, which undergo reverse transcription producing the cDNA templates for sequencing. Bulk PCR amplification was then performed to establish a library, which was for 3'end sequencing at last.