Figure 3.

VG16KRKP-mediated interference of the DNA-related processes in C. neoformans. (A) Confocal images of C. neoformans H4-mCherry incubated either without peptide (i) or with FITC-tagged VG16KRKP incubated for 8 h (ii, iii, iv) with ii showing the image through FITC channel; iii showing the image through mCherry channel; i, and iv showing the FITC/mCherry superimposed/merged images. ii, iii, and iv show the peptide to be membrane bound as well as localized in the nucleus with an observable enlargement of the nucleus in few cases, marked by the arrow, depicting nuclear fragmentation. (B) Plot of fluorescence intensity of ribo-green bound to RNA versus VG16KRKP concentration showing complete transcriptional inhibition in vitro, at 40 μM concentration. (C) Flow cytometry analysis showing the effect of peptide on the nuclear mass in C. neoformans. As shown, before treatment (left panel), cells are present in both interphase (G1 peak) and mitotic phase (G2/M peak). Peptide untreated cells also show the similar profile (central panel) while peptide treated cells (right panel) show a reduction in mitotic cell population and an emergence of dead cell peak (Sub G1). The cell populations from the same experiment were quantified and are shown in the table. To see this figure in color, go online.