Figure 2. Hist-5* retains antifungal activity and uptake similar to native Hist-5.

(A) C. albicans cells were pre-incubated with increasing concentrations of Hist-5 (blue) or Hist-5* (purple) for 1.5 h at 37 °C in PPB pH 7.4. Aliquots were resuspended in YPD and cell growth was measured by OD₆₀₀ after 48 h incubation at 30 °C. Data and error bars represent the average from three biological replicates. Differences in Hist-5 and Hist-5* OD₆₀₀ values were not statistically significant (p > 0.04, n = 3). (B) Timelapse microscopy images of cells treated with 12.5 μM Hist-5* at room temperature (RT) over 30 min in PPB. Corrected total cell fluorescence (CTCF) per cell is reported over time for each fluorophore channel. Scale bar = 10 μm. Error bars represent the standard deviation between three replicate experiments. (C) Confocal fluorescence microscopy images of cells treated with Hist-5* + vehicle control (purple), + spermidine (red), or + Hist-5 (blue) as competitive substrates for cellular uptake. Cells imaged at RT for 5 min in PPB. CTCF for individual fluorescence channels under each treatment condition were quantified, each dot representing fluorescence values from individual cells on experiments carried out on three separate days. Scale bar = 10 μm. (* indicates p < 0.05, ** indicates p < 0.01, n = 3)