Figure 7

GFP-Cys3 cellular localization and expression pattern in western blotting in mutant background. (a) Bars represent percentage of GFP and DAPI localization accessed by fluorescence microscopy in CNU080 (wild type), cna1Δ (CNU119), cnb1Δ (CNU121), and gpp2Δ (CNU125) cultivated for 2 h in YEPD and SD-N + Cys/Met; (b) fluorescence microscopy images showing localization of DAPI and GFP in YEPD (left) and SD-N + Cys/Met (right side). Merge is the overlay of DAPI and GFP images and localization is represented by the yellowish color; (c) western blotting of total proteins extracted from strain CNU080, CNU119, CNU121, and CNU125 cultivated for 1 and 2 hours in YEPD and SD-N + Cys/Met. Mouse Anti-GFP primary antibody was used at 1:2000 dilution and secondary anti-mouse horseradish peroxidase-linked antibody was applied at 1:2000 dilution. Loading control was done with histone H3 protein with rabbit anti-His3 antibody (1:2000) as primary and anti-rabbit secondary antibody horseradish peroxidase linked (1:2000). The original western blot images are shown in Supplementary data; (d) expression pattern by qPCR of C. neoformans sulfate permease (SUL1) in wild type (H99), gpp2Δ (CNU125), cna1Δ (CNU119), and cnb1Δ (CNU121) in YEPD. Asterisks denote the statistically significant differences encountered between wild type H99 and the mutants. *p < 0.05, ***p < 0.001.