Journal: Cell Death & Disease
Article Title: A broad-spectrum antibiotic, DCAP, reduces uropathogenic Escherichia coli infection and enhances vorinostat anticancer activity by modulating autophagy
doi: 10.1038/s41419-018-0786-4
Figure Lengend Snippet: 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)
Article Snippet: Human osteosarcoma cells U2OS, human embryonic kidney cells 293 (HEK293), human melanoma cancer cells A375 and human urinary bladder epithelial carcinoma cells (5637, HTB9) were acquired from American Type Culture Collection and the National Collection of Type Cultures (ATCC), were grown in Dulbecco’s modified Eagle’s medium (DMEM, Euroclone, ECB7501L) with 4.5 g/L glucose supplemented with 10% fetal bovine serum (FBS, Gibco, 10270106), 2 mM L-glutamine and 1% Penicillin/streptomycin.
Techniques: Bacteria, Concentration Assay, Western Blot, Control, Infection, Membrane, MANN-WHITNEY, Transfection
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