Figure 2
From: Enterobacter bugandensis: a novel enterobacterial species associated with severe clinical infection
Growth of EB-247 in human serum and physical characterization of EB-247 O-antigens and their role in cytokine induction in vitro. (A) The growth pattern of EB-247 in 50% human serum. Five µL from overnight cultures of individual strains were suspended in human serum and incubated at 37 °C for 22 h in a 96-well microtiter plate. The OD600 was measured in every hour by the incubator cum microtiter plate reader. Data represented means ± SD of measurements performed in triplicate. (B) Characterization of EB-247 lipopolysaccharide (LPS). The LPS of individual strains were extracted by using the hot phenol water method and electrophoresed in a Tris-Glycine polyacrylamide gel. Further, the gel was stained by using a conventional silver staining protocol. (C) Estimation of fold change in the mRNA expression of IL-1, IL-6 and TNF-α upon LPS stimulation in BMDM cells. The cells were stimulated with 0.5 µg/mL of LPS isolated from EB-247 and SB 300. The BMDM cells were harvested 4 h post stimulation and the cDNA was prepared from the isolated total RNA. In addition, quantitative gene expression was measured using qRT PCR. The data represents the mean with standard deviation of three individual experiments with three replicates of each.
