Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: CgTOG1 confers resistance to oxidative stress inducers but is not required for the utilization of alternative carbon sources . (a) Comparison of spot growth assays of the KUE100 C. glabrata wild type and derived Δcgtog1 deletion mutant, as well as the L5U1 C. glabrata wild type strain, harboring the pGREG576 cloning vector, or the pGREG576_MTI_ CgTOG1 expression plasmid, in the presence of glucose, lactate, glycerol or oleate as carbon sources. (b) Comparison of spot growth assays of the KUE100 C. glabrata wild type and derived Δcgtog1 deletion mutant, as well as the L5U1 C. glabrata wild type strain, harboring the pGREG576 cloning vector, or the pGREG576_MTI_ CgTOG1 expression plasmid, in the presence of oxidative stress inducers H 2 O 2 and menadione. (c) Comparison of spot growth assays of the KUE100 C. glabrata wild type and derived Δcgtog1 deletion mutant, as well as the L5U1 C. glabrata wild type strain, harboring the pGREG576 cloning vector, or the pGREG576_MTI_ CgTOG1 expression plasmid, in the presence of distinct carbon sources and the oxidative stress inducer H 2 O 2 . (d) Comparison of spot growth assays of the KUE100::URA- C. glabrata wild type strain and the derived KUE100_ Δcgtog1 :URA- deletion mutant, harboring the pGREG576 cloning vector, or the pGREG576_MTI_ CgTOG1 expression plasmid, in the presence of the oxidative stress inducer H 2 O 2 . The inocula were prepared as described in the materials and methods section. Cell suspensions used to prepare the spots were 1:5 (b) and 1:25 (c) dilutions of the cell suspension used in (a). The displayed images are representative of at least three independent experiments

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Derivative Assay, Mutagenesis, Clone Assay, Plasmid Preparation, Expressing

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: CgTOG1 contributes to reduce the intracellular accumulation of ROS . Comparison of intracellular ROS concentration of the KUE100 C. glabrata wild type and derived Δcgtog1 deletion mutant in control conditions or upon 1 h of H 2 O 2 stress. The estimation of intracellular ROS is based on the fluorescence intensity values exhibited by yeast cells upon incubation with the cell-permeant 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate indicator H 2 DCFDA. The displayed values correspond to at least three independent experiments. Error bars represent the corresponding standard deviation

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Concentration Assay, Derivative Assay, Mutagenesis, Fluorescence, Incubation, Standard Deviation

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: CgTOG1 is localized to the nucleus . Fluorescence of L5U1 C. glabrata cells harboring the pGREG576_MTI_ CgTOG1 plasmid after 5 h of copper-induced recombinant protein production in BM-U medium. (a,b) Cells were washed with PBS buffer and transferred to YP medium with glucose or oleate as carbon source for 1 h. (c,d) Cells were washed with PBS buffer and transferred to fresh medium with or without 15 mM H 2 O 2 for 1 h. Results indicate that CgTog1_GFP fusion protein localized to the nucleus under all conditions tested. The displayed images are representative of at least three independent experiments

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Fluorescence, Plasmid Preparation, Recombinant

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: CgCTA1 expression is independent of CgTOG1 . Comparison of the variation of CgCTA1 transcript levels determined by RT-PCR in the KUE100 C. glabrata wild type and derived Δcgtog1 deletion mutant in control conditions or upon 1 h of 15 mM H 2 O 2 stress. Transcript levels of CgACT1 were used for normalization. Expression values are the average of at least three independent experiments. Error bars represent the corresponding standard deviation

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Mutagenesis, Standard Deviation

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: C. glabrata gene expression during oxidative stress and its regulation by CgTOG1 . (a) Heatmap depicting the global transcriptomics response to H 2 O 2 in KUE100 C. glabrata wild type and the derived Δcgtog1 . (b) Systematic analysis of differentially expressed genes regulated by CgTOG1 with the help of enriched KEGG pathways calculated with FungiFun2. The significantly enriched pathways are shown

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Expressing, Derivative Assay

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: List of CgTog1 activated genes which are also upregulated in the wild type oxidative stress response caused by H 2 O 2 . Expression values represent log 2 fold changes

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Expressing

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: CgTOG1 is required for survival upon phagocytosis . (a) The concentration of viable KUE100 C. glabrata wild type (black bars) or derived Δcgtog1 deletion mutant (gray bars) after co-culture with G. mellonella hemocytes using a MOI of 1:5. (b) The concentration of viable L5U1 C. glabrata wild type strain, harboring the pGREG576 cloning vector (black bars), or the pGREG576_MTI_ CgTOG1 expression plasmid (gray bars) after co-culture with G. mellonella hemocytes using a MOI of 1:5. The displayed results are relative to the concentration of viable cells inoculated at time zero and are the average of at least six independent experiments. Error bars represent the corresponding standard deviation

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Concentration Assay, Derivative Assay, Mutagenesis, Co-Culture Assay, Clone Assay, Plasmid Preparation, Expressing, Standard Deviation

Journal: Virulence

Article Title: A new regulator in the crossroads of oxidative stress resistance and virulence in Candida glabrata : The transcription factor CgTog1

doi: 10.1080/21505594.2020.1839231

Figure Lengend Snippet: The CgTOG1 regulon in the transcriptomics context of macrophage engulfed cells . (a) Number of differentially expressed genes that constitute the CgTOG1 activated genes in either control conditions or H 2 O 2 stress, compared to the activated transcriptional response of C. glabrata upon macrophage engulfment. Overlapping genes were identified based on their systematic ORF designations. (b) Systematic analysis of genes commonly activated by CgTOG1 and upon macrophage engulfment, with the help of enriched FunCat functional categories calculated with FungiFun2. The significantly enriched pathways are shown

Article Snippet: The distribution of CgTog1_GFP fusion protein in C. glabrata living cells was detected by fluorescence microscopy in a Zeiss Axioplan microscope (Carl Zeiss MicroImaging), using excitation and emission wavelength of 395 and 509 nm (GFP) or 358 and 461 nm (DAPI), respectively.

Techniques: Functional Assay