To study the role of menaquinone in Salmonella yqiC virulence, 100 μg/mL menaquinone (Sigma-Aldrich) was added into the LB broth containing S. Typhimurium ΔyqiC and then coincubated at 37°C for 18 h. The total RNA of S. Typhimurium ΔyqiC was isolated and processed as described for the qRT-PCR analysis. The mRNA expression levels of fimA, fimZ, flhD, fliZ, invF, and sseB in the S. Typhimurium ΔyqiC complemented with menaquinone were compared with those in S. Typhimurium SL1344.
Menaquinone
Manufactured by Merck Group
Menaquinone is a type of vitamin K2 that plays a role in blood clotting and bone metabolism. It is a naturally occurring compound found in various food sources, such as fermented foods and some animal products. Menaquinone functions as an essential cofactor in the carboxylation of certain proteins involved in these physiological processes.
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2 protocols using menaquinone
1
Menaquinone's Role in Salmonella Virulence
2
Vascular Smooth Muscle Cell Calcification
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Human aortic vascular smooth muscle cells (HASMC) were obtained from Cambrex Bioscience (Wokingham, United Kingdom) and cultured in medium-199 (Sigma-Aldrich, St. Louis, MO) containing 15% fetal bovine serum (FBS), 2 mmol/L l-glutamine, 100 U/mL penicillin G, and 100 μg/mL streptomycin. Calcification of HASMC cultures was induced by the method of Wada et al. 14 Briefly, HASMC were cultured in normal growth medium for 4 days and then switched to medium containing vehicle control, high phosphate levels (final concentration 5 mM), warfarin (10 μM; Sigma-Aldrich, St. Louis, MO), or warfarin plus high phosphate levels for 10 days. The cell culture medium was replaced with fresh medium every other day. The antagonistic effect of vitamin K1 (phytomenadione, 5 μM; Sigma-Aldrich, St. Louis, MO) and vitamin K2 (menaquinone, 25 μM; Sigma-Aldrich, St. Louis, MO) on mineral deposition in HASMC induced by warfarin was also examined. The extent of transdifferentiation from HASMC into osteoblasts was evaluated using α-smooth muscle actin, osteocalcin, and Runx2 levels that were examined by immunoblotting as described below. Cell cultures were stained for mineral deposition using the von Kossa method as previously described. 14
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