Menadione

P. gingivalis W83 was grown in hemin (5 µg/mL) (Solarbio, Beijing, China) and menadione (1 µg/mL) (Solarbio, Beijing, China) containing brain heart infusion broth (BHI) (Oxiod, Basingstoke, UK) anaerobically (37 °C, 85% N2, 10% H2, 5% CO2). 10^{^9} CFU/mL bacteria were collected for following use.
For mice oral infection, the bacterial precipitates obtained after centrifugation were resuspended in a 2% carboxymethylcellulose (CMC) solution.
In the case of the bacteria-cell co-culture experiments, the bacterial precipitates obtained after centrifugation were resuspended in 10% fetal bovine serum (FBS)-containing DMEM and then co-cultured with cells at a multiplicity of infection (MOI) of 100. [22 , 23 (link)].

P. gingivalis W83 was grown in brain heart infusion broth (BHI) (Oxiod, Basingstoke, UK) enriched with hemin (5 g mL^-1) (Solarbio, Beijing, China) and menadione (1 g mL^-1) (Solarbio), in an anaerobic atmosphere (85% N₂, 10% H₂, 5% CO₂), at 37°C. Bacteria were collected when the culture reached an optical density at 600 nm of approximately 1.0 (~10⁹ CFU mL^-1) and were used for infection of unprimed BMDMφ at a multiplicity of infection (MOI) of 100, ^{30 (link)} in the presence or absence of 5 μM Stattic (Selleck, TX, USA), a nonpeptidic small molecule that selectively inhibits the phosphorylation of STAT3. P. gingivalis W83 were incubated with the unprimed BMDMφ for 12 h.

P. g ATCC 33,277 was cultured on blood agar mediums containing tryptic soy agar (Solarbio, China), supplemented with 1 µL/mL vitamin k3 (Landbridge Technology Co. Ltd., China) and 5 µL/mL hemin (Solarbio, China), in an anaerobic chamber under conditions of 85% N₂, 10% H₂, and 5% CO₂ at 37 °C for 7 days. The growth curves were evaluated to establish the log phase under a 600 nm optical density (OD) to evaluate the CFU/mL concentration of microorganisms. P. gingivalis colonies were transferred to a 15 mL tube containing brain-heart infusion (BHI), hemin and menadione (Solarbio, China) and cultured for 48 h under anaerobic conditions.