Bifidobacterium breve Enhances Growth and Alters the Transcriptome of Trichomonas vaginalis With Transient Modulation of Host Inflammatory Responses
T. vaginalis co-cultured with B. breve or T. vaginalis alone (negative control) were transferred into centrifuge tubes and centrifuged at 2500 rpm for 5 minutes. The supernatant was removed, and TRIzol reagent was added and mixed thoroughly. T. vaginalis only culture in serum-free YI-S and incubated anaerobically for four hours.  Total RNA was extracted using the Quick-RNA MiniPrep Kit (Zymo Research, Cat. No. R2052) according to the manufacturer's instructions. Briefly, cells were lysed in RNA Lysis Buffer, and the lysates were passed through spin columns for RNA binding. After on-column DNase I digestion to remove genomic DNA, the columns were washed and RNA was eluted in RNase-free water. RNA concentration and purity were determined using a NanoDrop spectrophotometer, and RNA integrity was verified with an Agilent Bioanalyzer. RNA-seq libraries were constructed using the Illumina Poly(A) Capture (Cat. No. 20777928), cDNA Synthesis (Cat. No. 20761354), and RNA Prep, Ligation (Cat. No. 20778789) kits according to the manufacturer's protocols.