Table 2 Enhancement of cell surface Lewis antigen expression by the growth of cultures in the presence of cholesterol.a   fold increase compared to parallel cholesterol-free CHIR-99021 culture   Lewis X Lewis Y   mean ± SEM (n) P value mean ± SEM (n) P value 26695 4.32 ± 0.36 (6)

0.0002 not done   SS1 not done   1.88 ± 0.08 (5) 0.0004 G27 wild type 2.85 ± 0.42 (8) 0.0033 2.22 ± 0.24 (8) 0.0016 G27 cgt::cat 3.69 ± 0.34 (5) 0.0013 2.88 ± 0.30 (5) 0.0034 G27 lpxE::cat 2.59 ± 0.50 (6) 0.025 2.47 ± 0.43 (7) 0.014 a Lewis antigens were quantitated in replicate whole-cell ELISA analyses of paired samples grown in the presence or absence of 50 μg/ml cholesterol. The antigen load was 300 ng cellular protein per well. Ratios for plus:minus cholesterol were calculated from duplicate net absorbance www.selleckchem.com/products/Imatinib-Mesylate.html readings

in each assay, and ratios determined in five to eight independent ELISA runs were then averaged. P values were calculated in two-tailed Student t-tests for the null hypothesis that the ratio equals 1. Table 3 Enhanced cell surface Lewis antigen expression CDK inhibitor is cholesterol-specific   fold increase compared to parallel cholesterol-free culture   Lewis X Lewis Y   mean ± SEM (n) P value mean ± SEM (n) P value cholesterol 2.96 ± 0.22 (5) .0008 2.48 ± 0.10 (4) .0007 β- sitosterol 1.80 ± 0.47 (4) 0.19 1.19 ± 0.13 (3) 0.28 taurocholate 0.64 ± 0.16 (4) 0.12 0.84 ± 0.20 (3) 0.52 Lewis antigens were quantitated in replicate whole-cell ELISA analyses of pairwise cultures of H. pylori G27 grown in the presence or absence of 130 μM cholesterol, or an equal concentration

of β-sitosterol or sodium taurocholate. The antigen load was 300 ng cellular protein per well. Ratios for plus:minus cholesterol were calculated from duplicate net absorbance readings in each assay, and ratios determined in three to five independent ELISA runs were then averaged. P values were calculated in two-tailed Student t-tests for the null hypothesis that the ratio equals 1. Figure 4 Growth in cholesterol specifically enhances cell surface display of Lewis antigens. Whole cell Anidulafungin (LY303366) ELISA assays were performed on samples of H. pylori strain 26695 (upper left), SS1 (lower left), or G27 (upper and lower right). Parallel cultures were grown overnight in defined medium containing 130 μM of the following additions: circles, no addition; squares, cholesterol; triangles, β-sitosterol; X, taurocholate. Varying amounts of cell suspension corresponding to known amounts of cellular protein were applied to duplicate wells of ELISA plates, and immunoassayed for the presence of Lewis X or Lewis Y antigen as described in Methods. Negative control samples of E. coli HB101, or buffer-only blanks, fell on the dotted line. Absorbance readings for individual wells are plotted.