Journal: mBio

Article Title: Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids

doi: 10.1128/mBio.00647-15

Figure Lengend Snippet: Measurements of sphingolipids upon treatment with BHBM. (a) Thin-layer chromatography analysis of sphingolipids (see diagram) isolated from untreated or BHBM-treated C. neoformans cells after in vivo labeling with [ 3 H]palmitate ( 3 H-Palm). (b to h) Lipid analysis by LC-MS. (b) C18 hydroxy (Δ8) 9 methyl-glucosylceramide (GlcCer); (c) C18 dihydroceramide (C18 dhCer); (d) sphingosine and dihydrosphingosine (SPHs); (e) sphingosine-1-phosphate and dihydrosphingosine-1-phosphate (SPH-1-P); (f) C18 hydroxyceramide (C18 OH-Cer); (g) C18 hydroxy Δ8-ceramide (C18 OH-Δ8-Cer); and (h) C18 hydroxy Δ8, 9 methyl-ceramide (C18 OH-Δ8, 9Me-Cer). Values that are significantly different are indicated as follows: *, P < 0.05 comparing treated to untreated (no drug). Statistical analysis was performed using analysis of variance (ANOVA) test. Statistical significance is accepted at a P value of <0.05. Data were compiled from three independent experiments.

Article Snippet: Fluorescence intensity of yeasts labeled with NBD C6-ceramide were compared using a FACScan flow cytometer (BD Biosciences, Franklin Lakes, NJ).

Techniques: Thin Layer Chromatography, Isolation, In Vivo, Labeling, Liquid Chromatography with Mass Spectroscopy

Journal: mBio

Article Title: Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids

doi: 10.1128/mBio.00647-15

Figure Lengend Snippet: Effect of BHBM on Golgi morphology in C. neoformans and yeast-to-hyphal differentiation in C. albicans . (A) Control or BHBM-treated cells were stained with NBD C6-ceramide (Golgi and Golgi-derived compartments, green fluorescence). DIC, differential interference contrast; NBD-Cer, NBD 6-ceramide. Bars, 5 µm. (B) FACS analysis shows NBD C6-ceramide accumulation in C. neoformans . The gray histogram corresponds to C. neoformans yeast in the absence of NBD C6-ceramide. CT, control (no BHBM treatment). (C) Percentage of yeast and hyphal forms under control conditions and after BHBM treatment. Statistical analysis was performed using analysis of variance (ANOVA) test. Statistical significance is accepted at a P value of <0.05. The images in panels A and B are representative of three separate experiments. The data in panel C were compiled from three independent experiments.

Article Snippet: Fluorescence intensity of yeasts labeled with NBD C6-ceramide were compared using a FACScan flow cytometer (BD Biosciences, Franklin Lakes, NJ).

Techniques: Staining, Derivative Assay, Fluorescence