Nystatin nyst

We calculated the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of the twelve clinical strains of C. parapsilosis using Fluconazole (FLZ; Sigma-Aldrich, St. Louis, MO, USA), Amphotericin B (AmB; Sigma-Aldrich, USA), Nystatin (NYST; Sigma-Aldrich, USA), and Caspofungin (CAS; Sigma-Aldrich, USA). A broth microdilution assay was performed under the guidelines recommended in the standard M27 document (4th ed.) presented by the Clinical and Laboratory Standard Institute [14 ]. The stock solution of the prior mentioned drugs was prepared using dimethyl sulfoxide (DMSO; Merck, RSA) and the test concentration ranged from 1250 to 9.77 µg/mL for FLZ, 62.5 to 0.49 µg/mL for AmB, 250 to 19.5 µg/mL for NYST, and 8 to 0.06 µg/mL for CAS. Sterility and growth control were included for comparison. The plates were incubated at 37 °C for 48 h. The MIC results were determined by visual observation, where antifungal drugs with the lowest concentration inhibited the fungal growth when compared to the growth control was considered as MIC value.
For estimating MFC, 10 µL from the wells with different concentrations of antifungal drugs were subcultured onto Sabouraud dextrose agar (SDA; Merck, RSA) plates followed by incubation at 37 °C for 48 h. The MFC was determined by observing the lowest concentration with no growth on the agar plate [15 (link)].

RAW264.7 macrophages, cells were preincubated with chlorpromazine (CHL; 50 μM, Sigma-Aldrich) to inhibit clathrin-mediated endocytosis, nystatin (NYST; 10 μg/mL, Sigma-Aldrich) to inhibit caveolae-mediated endocytosis, and amiloride (AMI; 100 μM, Sigma-Aldrich) to inhibit macropinocytosis. After 30 min at 37 °C, the inhibitor solutions were removed, and freshly prepared c6-loaded KT NPs in medium (NPs:DMEM (1:5, v/v)) were added (0.25 mg/mL) and further incubated for another 8 h. Subsequently, the cells were washed and lysed as previously described. The groups treated with c6-loaded NPs but without inhibitor at 4 °C or at 37 °, were used as negative and positive control, respectively. The percentage uptake after treatment with the inhibitors was normalized to the positive control uptake, which was expressed as 100% (Equation (3)).
$\%Uptake=\frac{\left[c6\right]inhibitor}{\left[c6\right]control}x100$