The C. albicans SC5314 strain was grown overnight as described above, washed with phosphate-buffered saline (PBS), used to inoculate YPD containing 10% fetal bovine serum (FBS) at a 1:30 dilution, and incubated at 37°C for 90 min in the presence or absence of compound 9029936 at 5 μM as previously described (30 (link)). RNA was extracted by using a hot-acid-phenol protocol (78 ). Three biological replicates were obtained for each condition (treated and untreated). To determine the final RNA concentration and quality, samples were analyzed using a 2100 series bioanalyzer (Agilent Technologies, CA).
RNA sequencing was performed at the Genome Sequencing Facility at the Greehey Children’s Cancer Research Institute at the University of Texas Health Science Center at San Antonio. Briefly, cDNA libraries for RNA-seq analysis were prepared from total RNA samples using an Illumina TruSeq stranded mRNA-seq kit. RNA sequencing was performed using an Illumina HiSeq 2000 machine (San Diego, CA) to obtain 100-bp paired-end reads. After the sequencing run, demultiplexing with CASAVA was employed to generate a fastq file for each sample.
RNA sequencing was performed at the Genome Sequencing Facility at the Greehey Children’s Cancer Research Institute at the University of Texas Health Science Center at San Antonio. Briefly, cDNA libraries for RNA-seq analysis were prepared from total RNA samples using an Illumina TruSeq stranded mRNA-seq kit. RNA sequencing was performed using an Illumina HiSeq 2000 machine (San Diego, CA) to obtain 100-bp paired-end reads. After the sequencing run, demultiplexing with CASAVA was employed to generate a fastq file for each sample.
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