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. 2021 Feb 15;131(4):e140333. doi: 10.1172/JCI140333

Figure 8. In vivo validation of nlpD complementation and treatment.

Figure 8

Mice were infected with E. coli 83972, SN25, or SN25-pRH320 and sacrificed after 24 hours. (A) NlpD-rpoS reconstitution increased Pol II inhibition in vivo. Tissue sections were stained with anti–Pol II-p antibodies and the fluorescence intensity was quantified (mean ± SEM). Reduced Pol II-p staining in the bladder mucosa of mice infected with E. coli 83972 or E. coli SN25-pRH320 (P = 0.006) compared with E. coli SN25. (B) Neutrophil (left) and bacterial counts (middle) in urine, and bladder pathology scores (right) were reduced in mice infected with E. coli SN25-pRH320 compared with E. coli SN25 (PMNs, polymorphonuclear leukocytes; CFU, colony forming unit). *P < 0.05, **P < 0.01, ***P < 0.001 compared with E. coli 83972 by Kruskal-Wallis test with Dunn’s multiple-comparison test. (C) E. coli SN25–infected, NlpD-treated mice showed lower neutrophil counts (left), bacterial counts (middle), and bladder pathology scores (right) than untreated mice (24 hours). ***P < 0.001 by Mann-Whitney test. Urine PMNs and CFUs for SN25-infected mice are presented in B and C. Scale bar: 50 μm. Data are presented as mean ± SEM (n = 5 mice, 2 independent experiments).