Summary of BHT inhibition and chemical epistasis data.

<p>In the Spider, 10% serum, Lee's, and GlcNAc columns, “+” refers to statistically significant BHT inhibitory activity (<i>P</i> value of <0.05 compared to the DMSO control; see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025395#pone-0025395-g002" target="_blank">Figure 2</a>), whereas “−” refers to no statistically significant BHT inhibitory activity (<i>P</i> value of >0.05 compared to the DMSO control). In the <i>efg1</i>-T206E, <i>ADH1pr-CPH1</i>, db-cAMP, <i>ras1</i>-G13V, and <i>gpa2</i>-Q355L columns, “+” refers to the constitutively active mutant or db-cAMP being epistatic to the added BHT inhibitor, whereas “−” refers to the added BHT inhibitor being epistatic to the constitutively active mutant or db-cAMP. NT, not tested.</p

Chemical epistasis with <i>efg1</i>-T206E mutant and <i>ADH1pr-CPH1</i> overexpression.

<p><b>A,</b> Wild-type strain SC5314 (white bars) and <i>efg1</i>-T206E mutant strain AV55 (black bars) were grown in Spider media with the indicated BHT inhibitors (100 µM final concentration) for 6 h at 37°C prior to quantification of morphological phenotype as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025395#pone-0025395-g002" target="_blank">Figure 2</a>. Asterisks indicates a <i>P</i> value of <0.05 compared to the corresponding wild-type SC5314 plus BHT inhibitor control. <b>B, </b><i>ADH1pr-CPH1</i> strain CDH72-1 was assayed in the presence of the indicated BHT inhibitors as in A. Asterisks indicates a <i>P</i> value of <0.05 compared to the wild-type SC5314 plus BHT inhibitor control.</p

BHT inhibition in different hyphal-inducing media.

<p>Wild-type strain SC5314 was grown in the four different hyphal-inducing media for 6 h at 37°C. At least 100 cells were counted for each sample in duplicate and all assays were repeated at least twice. Standard deviations are shown for each sample. Asterisks indicates a <i>P</i> value of <0.05 compared to the DMSO control.</p

Chemical epistasis with cAMP and <i>ras1</i>-G13V mutant.

<p><b>A,</b> Wild-type strain SC5314 was incubated in the absence (white bars) and presence (black bars) of 10 mM <i>N</i>⁶,2′-<i>O</i>-dibutyryl-cAMP (db-cAMP) for 6 h at 37°C. Quantification of morphological phenotypes was as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025395#pone-0025395-g002" target="_blank">Figure 2</a>. Asterisks indicates a <i>P</i> value of <0.05 compared to the corresponding –db-cAMP control. <b>B,</b> The <i>ras1</i>-G13V constitutively active mutant strain DH409 was incubated in Spider media with the indicated BHT inhibitors for 6 h at 37°C as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025395#pone-0025395-g003" target="_blank">Figure 3</a>. Asterisks indicates a <i>P</i> value of <0.05 compared to the corresponding wild-type SC5314 plus BHT inhibitor control.</p

Chemical epistasis with <i>gpa2</i>-Q355L mutant and <i>ADH1pr-GPR1</i> overexpression.

<p><b>A,</b> Wild-type strain SC5314 (white bars) and <i>gpa2</i>-Q355L mutant strain HTC7 (black bars) were grown in Spider media with the indicated BHT inhibitors (100 µM final concentration) for 6 h at 37°C prior to quantification of morphological phenotype as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025395#pone-0025395-g002" target="_blank">Figure 2</a>. Asterisks indicates a <i>P</i> value of <0.05 compared to corresponding wild-type SC5314 plus BHT inhibitor control. <b>B,</b> The <i>ADH1pr-GPR1</i> strain YTC014 was spotted onto Spider solid media containing either DMSO or clozapine (∼100 µM final concentration) and incubated for 7 days at 37°C.</p

BHT pathways and inhibitors.

<p><b>A,</b> Morphological signaling pathways in <i>C. albicans</i>. For simplicity, only the Efg1, Cph1, Cph2/Tec1, and Rim101 pathways are shown. <b>B,</b> BHT inhibitors used in this study.</p