Figure Caption/Comments:

Fig. 6

Ezetimibe is protective in a zebrafish model of S. Typhi infection. (A) Fish infected with S. Typhi prgH had increased survival compared with fish infected with WT S. Typhi (P = 0.01). The survival curve was carried out for 5 d; fish were checked once each day. (B) Zebrafish were scored 24 h post S. Typhi infection as cleared (no bacteria), localized (bacteria only in the swim bladder), disseminated (bacteria found outside the swim bladder), or dead (fish dead due to bacterial burden). The swim bladders are denoted by red circles; bacteria are denoted by the red arrows. (C) Fish infected with the S. Typhi prgH mutant had increased clearance of bacteria at 24 h (P = 0.03). (D) Ezetimibe had no effect on S. Typhi bacterial growth. Bacteria were diluted from an overnight stock and grown with DMSO or 10 µM ezetimibe. The OD600 was taken every 30 min for 3.5 h. Data points are the mean from two separate experiments. (E) Ezetimibe decreased filipin staining in fish. Twenty-four–hour pretreatment with 10 µM ezetimibe reduced filipin (0.05 mg/mL) staining (P = 0.003) in zebrafish; n = 20 fish from two separate experiments; P value from an unpaired t test. (F) Fish pretreated with ezetimibe had increased survival from S. Typhi infection compared with DMSO-pretreated controls (P = 0.03). (G) Ezetimibe treatment increased bacterial clearance in fish. Twenty-four–hour pretreatment with 10 µM ezetimibe increased the percentage of fish that cleared the bacteria 24 h postinfection from 8 to 30% (P = 0.009). Infection data for each survival curve and clearance comparisons are from three independent experiments with a minimum of n = 60 fish. P values from survival curves are from the Mantel–Cox test; P values for other comparisons are from unpaired t tests.

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