Fig. 6. The combination of BRI+CAS is not toxic to human cells and can significantly decrease the A. fumigatus fungal burden in a chemotherapeutic murine model.

a A549 lung cells grown in the absence or presence of different concentrations of BRI and CAS. DMSO 10% was used as positive control. Percentage of cell viability is expressed as the absorbance value of experiment well/absorbance value of control well x 100. All the results are the average of three repetitions ±SD. b A549 cells were seeded at a density of 106 cells/ml and challenged with A. fumigatus conidia at a multiplicity of infection of 1:10 in the absence or presence of different concentrations of BRI and CAS. After 24 h of incubation media was removed and the cell suspension were seeded in Sabouraud Dextrose Agar Media. The numbers of CFUs were determined after 24 h of growth. The CFU percentage for each sample was calculated and the results were plotted using Graphpad Prism (GraphPad Software, Inc., La Jolla, CA, USA). A p-value ≤ 0.001 was considered significant. c Quantification of the fungal burden in the infected lungs was determined 72 h post-infection. The total genomic DNA was extracted from the lungs, and fungal burden was determined by qPCR based on 18 S rRNA gene of A. fumigatus and an intronic region of the mouse GAPDH gene through the ratio between number of copies of the 18 S and number of copies of GAPDH. Data represent the means (±standard deviation) of two biological replicates (n = 10, representative of two independent experiments). P-values were calculated using One-way ANOVA with Tukey’s multiple comparisons test. **p < 0.0001.