Fig. 5: DCAP reduces autophagy-mediated uropathogenic E.coli infection.

a Growth curves of free uropathogenic E. coli bacteria (UPEC) in presence or absence of the indicated DCAP concentrations. b Measurements of the minimum inhibitory concentration (MIC) of growth against UPEC bacteria of DCAP, CQ or the well-characterized beta-lactam antibiotic, ampicillin. c Immunoblot analysis of protein samples from Bladder Epithelial Cells (BECs) treated 24 h with vehicle (DMSO) or 10 µM DCAP, showing DCAP-mediated accumulation of LC3-II and SQSTM1. GAPDH was used as a loading control. d The potential cytotoxicity of 10 µM DCAP against BECs was evaluated by lactate dehydrogenase (LDH) cytotoxic assay. Indicating the tolerability of BECs toward this dose of compound, cells treated for 24 h showed negligible differences in the release of LDH in the medium, compared with vehicle-treated cells. Reported as percentage of LDH cytotoxicity ± s.e.m., n = 6. e BECs were infected with a pathogenic UTI89 E. coli strain. After the removal of extracellular bacteria by gentamycin, an antibiotic that is not permeable through the human cell membrane, BECs were treated with 10 µM DCAP or vehicle for 24 h. The counting of intracellular bacteria from DMSO- and DCAP-treated infected cells is reported as an average of the colony-forming unit (CFU)/ml ± s.e.m., n = 12. **P < 0.01 DCAP vs. DMSO (Mann–Whitney test). f Immunoblot analysis of protein extract from BECs transfected with a siRNA sequences against ATG16L1 or a non-targeting element (control). GAPDH was used as a loading control. g Quantification of intracellular UPEC bacteria in infected siATG16L1 and control BECs treated with 10 µM or DMSO. Shown as average CFU/ml ± s.e.m., n = 7. *P < 0.05 DCAP vs. DMSO (Mann–Whitney test)