The activity of Cdc35 is indirectly inhibited by RAB.

<p>(A) YEM30 cells were subject to the indicated RAB treatment for 8 h and then were broken by beads beating. The lysates were assayed for the activity of Cdc35 by detecting the cAMP production as described in METHODS. (B) The purified catalytic domain of <i>C. albicans</i> Cdc35 was incubated with the indicated concentration of RAB for 20 min, then cAMP production was measured to determine the activity of Cdc35 under the treatment of RAB.</p

The morphology of <i>C. albicans</i> YEM30 challenged by RAB at the indicated conditions.

<p>The bar: 20 µm.</p

The antifungal efficacy of RAB <i>in vitro</i> and <i>in vivo</i> with FLC as positive control.

a<p>MIC₈₀<i>in vitro</i> has been reported <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041624#pone.0041624-Chang1" target="_blank">[41]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041624#pone.0041624-Otzen1" target="_blank">[42]</a>.</p>b<p>MIC₈₀ of FLC <i>in vitro</i> has been reported <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041624#pone.0041624-Larsen1" target="_blank">[43]</a>.</p>c<p>MIC₈₀<i>in vitro</i> has been reported in our lab <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041624#pone.0041624-Sun1" target="_blank">[10]</a>.</p

Farnesol and Dpp3 are stimulated by RAB in a dose-dependent manner.

<p>(A) <i>C. albicans</i> YEM30 was cultured in RPMI1640 medium containing RAB for 12 h, and farnesol in the supernatants was extracted and quantified by GC-MS. (B) The comparison of farnesol secretion of YEM30 strain at the indicated time between the vehicle group and RAB-treated one (10 µg/ml). (C) The expression of Dpp3 induced by RAB was observed using <i>DPP3-GFP-</i>BWP17 strain by confocal microscopy.</p

RAB confers improved nematodes’ survival by retarding the formation of invasive hyphae.

<p>(A) Nematodes were infected with <i>C. albicans</i> YEM30 for 2 h and then moved to pathogen-free liquid media in the presence of PBS (negative control), RAB (4 or 16 µg/ml) or 2 µg/ml of FLC (positive control). Each day the worms were monitored and the survival rate was calculated. <i>p</i><0.001 for 4RAB, 16RAB and FLC treated groups compared to PBS-treated group. (B) Nematodes were infected with the indicated <i>C. albicans</i> strains and then treated by a series of doses of RAB. After 5 days of therapy, the survival ratio in each group was calculated. (C) The nematodes were infected by <i>C. albicans</i> strain CASA1 with <i>GFP</i> tagged <i>CDR1</i> for 2 h and then moved into pathogen-free liquid media containing drugs or the vehicle for 5 days. And then they were observed in 4× objective magnification. Most nematodes in control group did not demonstrate any movement and developed filaments outside the body. However, the majority of the drug-treated were alive and not invaded by the hyphae (scale bars: 200 µm). Insert shows the images of nematodes captured at 200× magnification (scale bars: 100 µm). The representative worm was selected for confocal microscopic observation. Green fluorescence of hyphae cells and yeast cells was seen within the intestine of the dead nematode under the vehicle treatment. And no <i>C. albicans</i> cells were observed in the live worm challenged by RAB (about 60 nematodes in each group). (D) Enlarged view of the rectangle frame.</p

Organocatalytic Asymmetric C–H Vinylation and Arylation of <i>N</i>‑Acyl Tetrahydroisoquinolines

The first organocatalytic enantioselective oxidative C–H functionalization of <i>N</i>-acyl tetrahydroisoquinolines with vinyl and aryl boronates promoted by a chiral Brønsted acid is described. This metal-free process tolerates a wide range of electronically varied <i>N</i>-acyl tetrahydroisoquinolines and structurally diverse boronates with good to excellent enantioselectivities

Reduction of cAMP confers the defect of hyphae formation induced by RAB.

<p>(A) The expressions of hyphae-specific genes in response to RAB treatment. <i>C. albicans</i> YEM30 was cultured in RPMI1640 medium with or without 10 µg/ml RAB for 8 h at 37°C and then cells were harvested for RNA extraction. Genes involving in modulating hyphae formation or hyphae-specific were detected for transcript levels using qPCR. The expression of detected genes was normalized to <i>GSP1</i> and relative to the control group using formula 2^−ΔΔCT. The values are means ± standard deviations of three independent experiments. Asterisk (*) represents significance with <i>p</i><0.05. (B–C) Reduced intracellular cAMP level by RAB. YEM30 cells were cultured in RPMI1640 medium containing a series of RAB or the vehicle for 8 h (B) or 16 h (C) at 37°C. Cells were harvested for cAMP assay. (D) Exogenous cAMP restores the RAB-inhibited hyphae formation. YEM 30 cells were grown in RPMI1640 medium with the indicated treatment. After 3 h, the morphology was visualized by microscopy (the scale bar: 20 µm).</p

Gene-specific primers used for qPCR.

<p>Gene-specific primers used for qPCR.</p

Phase-contrast micrographs of BWP-<i>DPP3-GFP</i> cells challenged by bisbibenzyls compounds within 12 hours.

<p>A) <i>Candida albicans</i> cells grown in RPMI 1640 broth medium was captured using an Olympus fluorescent microscope. The results showed that cells co-incubated with A1 (16 µg/ml) displayed yeast cells, cells with no drugs formed mycelium after three hours and maintained substantial growth of hyphae. B) Cells grown on Spider solid media were photographed from 2 d to 5 d using 4× objectives of an Olympus fluorescent microscope.</p

Biofilm formed of BWP17-<i>DPP3-GFP</i> cells pretreated with bisbibenzyls compounds.

<p>A) Quantitative measurement of inhibition of biofilm formation with Microplate Reader. B) Confocal micrographs of BWP17-<i>DPP3-GFP</i> cells cultured in RPMI 1640 for 48 hours. C) Cells were stained with alamar blue and cultured for 4–6 hours in dark, the supernatant would become pink when cells grow into biofilm, otherwise turn into blue. The test was performed in quadruplicate. A11 represents the compounds that biofilm inhibitory activity >64 µg/ml, bk represents blank control and nk represents negative control.</p