Fig. 7

Glycine-dependent mechanism for HtrE, NfrA, YhcD expression. a Relative ATP abundance of E. coli K12 in the presence or absence of 100 mM glycine for the indicated length of time (n = 3). b Percent survival of E. coli K12 in the presence or absence of 5 mM ATP, 100 mM glycine or/and 100 µL serum (n = 3). c Relative cAMP abundance of E. coli K12 in the presence or absence of 100 mM glycine or/and 100 µL serum (n = 3). d Percent survival of the indicated mutants in the presence of 100 mM glycine, 100 µL serum or both (n = 3). e Western blot of CRP in E. coli K12 exposed to 0–5 mM ATP or 0–4 mM AMP. f Effect of 100 mM glycine, 100 µL serum or both on CRP expression. g Western blot of HtrE, NfrA, YhcD in these mutants. h qRT-PCR of htrE, nfrA, yhcD in these mutants (n = 3). i Flow cytometry quantification of anti-C9 neoantigen on outer membrane surface of the indicated mutants (n = 3). j Percent survival of the indicated mutants in the presence of 100 mM glycine, 100 µL serum or both (n = 3). k Western blot of HtrE, NfrA, YhcD of E. coli K12 in the presence of 100 mM glycine and the indicated amount of serum. l, m Western blot of HtrE, NfrA, YhcD of E. coli K12 (l) and Y17 (m) in the presence of 100 µL serum and the indicated concentrations of glycine. n Flow cytometry quantification of anti-C3b and anti-C9 neoantigen on E. coli K12 and Y17 in the presence or absence of 100 µL serum or 100 mM glycine or both. o, MST for the interaction of complement C3 with HtrE, NfrA, YhcD (n = 3). i, n 50,000 cells were record with forwarding versus side scatter and were gated before data acquisition. Results in (a–d, h–j, n, o) are displayed as mean ± SEM, and significant differences are identified (*p < 0.05, **p < 0.01) as determined by two-tailed Student’s t test. See also Supplementary Figs. 10–15