5810r unit

Blood was sampled from mice at days 0 and 7 of the experiment with a 29G insulin syringe (BD Biosciences (Franklin Lakes, NJ, USA)) by retrobulbar puncture. A sample of 150 μl was drawn during ischemia induction (day 0) and a sample of 300 μl was drawn prior to euthanasia at day 7. Blood samples were stabilized by EDTA and centrifuged (5,000 g, 10 min) in a tabletop 5810R unit (Eppendorf (Hamburg, Germany)) to obtain plasma for ELISA.

L. plantarum B21 cultures stored at −80°C were transferred to MRS agar (Oxoid) plates and incubated for 24 h at 37°C. For each experiment three colonies were selected at random and transferred to one of the four different broth media described in section Preparation of Growth Media and incubated for 24 h at 37°C. A total of 0.1 g of bacterial pellet per broth type was then collected from each experiment by centrifugation at 8,000 × g for 10 min using an Eppendorf 5810R unit (Eppendorf, Hamburg, Germany). These pellets were then collected in eppendorf tubes (Sigma Aldrich) and each was washed three times with milliQ H₂O for 5 min. These pellets were then used for metabolomics analysis via NMR and GC-MS. The viable cell count (CFU/mL) was ~10⁹⁻¹⁰ (for the unstressed control and Tween 80 stressed), ~10⁸ (glucose stressed), ~10⁷ (double-Tween 80 and glucose-stressed) as per Parlindungan et al. (2019 (link)). Each experiment was repeated three times (with three colonies) to make an n of nine for each treatment.