Journal: Frontiers in Microbiology

Article Title: SLA1 governs azole resistance in Candidozyma auris via mitochondrial ROS-driven efflux pump expression

doi: 10.3389/fmicb.2026.1948008

Figure Lengend Snippet: SLA1 deletion drives FLC resistance through upregulation of CDR1. (A) Spotting assays of WT and sla1Δ mutants. Strains were spotted onto YPD agar supplemented with the FLC, KTC, AMB and the plates were incubated at 30 for 2 days. (B) RT-qPCR analysis of ERG genes, CDR1 , CDR2 , MDR1 and PMC1 in WT and sla1Δ strains ( n = 3). (C) Spotting assays of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. Indicated strains were spotted with or without FLC. (D) FLC IC₅₀ values of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. (E) Correlation analysis between SLA1 and CDR1 transcript levels in 17 clinical C. auris isolates. (F) RT-qPCR analysis of CDR1 regulators CAP1 , TAC1 , and GSK3 in WT and sla1Δ strains ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Article Snippet: Drug-resistance tests were performed using 20 μg/mL FLC (MedChemExpress, cat# HY-B0101), 0.2 μg/mL ketoconazole (KTC) (MedChemExpress, cat# HY-B0105), 1.0 μg/mL Amphotericin B (AMB) (MedChemExpress, cat# HY-B0221), 10 mM NAC (Sigma, cat# 1009005), 20 μM Antimycin A (Sigma, cat# 77332), 50 μM Mito-TEMPO (MedChemExpress, cat# HY-112879), 60 μM Rotenone (MedChemExpress, cat# HY-B1756), and 1 mM H2O2 (Sigma, cat# 108600).

Techniques: Incubation, Quantitative RT-PCR, Analysis, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: SLA1 governs azole resistance in Candidozyma auris via mitochondrial ROS-driven efflux pump expression

doi: 10.3389/fmicb.2026.1948008

Figure Lengend Snippet: SLA1 deletion disrupts redox homeostasis and mitochondrial energy metabolism to regulate FLC resistance in C. auris . (A) GO analyses. GO term enrichment analyses were performed using the RNA-seq data from WT and sla1Δ strains. (B,C) Gene Set Enrichment Analysis (GSEA). Comparing the transcriptome data from the WT and sla1Δ strains. (D) Spotting assays of WT and sla1Δ strains. Strains were spotted onto YPD agar supplemented FLC, FLC + NAC, or FLC + H₂O₂ conditions. (E) Flow cytometry analysis of intracellular ROS levels in WT, and sla1Δ strains. Wild-type and sla1Δ cells were cultured YPD, loaded with MitoSOX, and then analyzed by flow cytometry. (F) Analysis of ROS levels. WT and sla1Δ strains were incubated at 30 °C for 12 h, then ROS levels were measured ( n = 4). (G) Analysis of ATP levels. WT and sla1Δ strains were incubated at 30 °C, then ATP levels were measured ( n = 3). (H) RT-qPCR analysis of genes involved in mitochondrial respiration and energy metabolism in WT and sla1Δ strains ( n = 3). (I) Analysis of ATP and ROS levels. Measurement of relative intracellular ATP and ROS content in laboratory wild-type susceptible strain and five clinical fluconazole-resistant C. auris isolates. ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Article Snippet: Drug-resistance tests were performed using 20 μg/mL FLC (MedChemExpress, cat# HY-B0101), 0.2 μg/mL ketoconazole (KTC) (MedChemExpress, cat# HY-B0105), 1.0 μg/mL Amphotericin B (AMB) (MedChemExpress, cat# HY-B0221), 10 mM NAC (Sigma, cat# 1009005), 20 μM Antimycin A (Sigma, cat# 77332), 50 μM Mito-TEMPO (MedChemExpress, cat# HY-112879), 60 μM Rotenone (MedChemExpress, cat# HY-B1756), and 1 mM H2O2 (Sigma, cat# 108600).

Techniques: RNA Sequencing, Analysis, Flow Cytometry, Cell Culture, Incubation, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: MIC values of fluconazole against Candida albicans .

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Determination of the fluconazole susceptibility phenotype of C. albicans by spot assay. C. albicans suspensions were adjusted to 1 × 10⁵ CFU/mL, followed by three successive 10-fold serial dilutions. The undiluted suspensions and serial dilutions were spotted from left to right onto YPD agar plates containing fluconazole (8 μg/mL) and incubated at 35 °C for 48 h.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques: Spot Test, Incubation

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Fluconazole MICs of C. albicans isolates prior to and following L. paragasseri treatment.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Lactobacillus paragasseri enhances the inhibitory effect of fluconazole on clinical isolates of C. albicans. (A–D) Representative spot-assay results for four clinical isolates of Candida albicans grown on solid YPD medium supplemented with 8 μg/mL fluconazole. The upper row shows the untreated C. albicans control group, whereas the lower row shows the group co-cultured with L. paragasseri . The initial concentration of the fungal suspension was 1 × 10⁵ CFU/mL, followed by three consecutive 10-fold serial dilutions spotted from left to right. Growth was assessed after 48 h of incubation. The images shown are representative of three independent experiments.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques: Spot Test, Control, Cell Culture, Concentration Assay, Suspension, Incubation

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Effect of L. paragasseri on biofilm formation of four clinical isolates of fluconazole-resistant C. albicans . (A–D) Quantitative analysis of biofilm biomass in four C. albicans isolates before and after treatment with L. paragasseri . According to the calculated OD values, the percentages of biofilm reduction for the above four tested isolates were 66, 28, 22, and 47%, respectively. Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined by unpaired two-tailed Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001. (E) After treatment with L. paragasseri , the hyphae of the four C. albicans isolates appeared shorter and less densely distributed, suggesting a reduction in biofilm formation. Scale bar, 10 μm. The images are representative of three independent experiments.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques: Analysis, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Effects of L. paragasseri on the expression of drug efflux pump-related genes of two clinical isolates of fluconazole-resistant C. albicans . (A–C) Following treatment with L. paragasseri , the expression levels of the drug efflux pump-related genes CDR1, CDR2 , and MDR1 were assessed in the LAB-V-24-112 isolate. In the LAB-V-24-112 isolate, L. paragasseri treatment reduced CDR1 expression by 1.5-fold, CDR2 by 1.2-fold, and MDR1 by 1.1-fold relative to untreated controls. (D–F) Following treatment with L. paragasseri , the expression levels of CDR1, CDR2, and MDR1 were assessed in the LAB-V-24-130 isolate. In the LAB-V-24-130 isolate, L. paragasseri treatment reduced CDR1 expression by 3-fold, CDR2 by 1.4-fold, and MDR1 by 1.4-fold relative to untreated controls. The results indicate that L. paragasseri treatment reduces the expression of these drug efflux pump-related genes in C. albicans . Data are presented as mean ± SD from three independent biological replicates. Statistical significance was determined by unpaired two-tailed Student’s t -test. * p < 0.05, *** p < 0.001.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques: Expressing, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Vaginal Lactobacillus paragasseri may attenuate fluconazole-resistant phenotypes in the subset of Candida albicans isolates and shows potential as a probiotic adjunct in antifungal therapy

doi: 10.3389/fmicb.2026.1951876

Figure Lengend Snippet: Lactobacillus paragasseri alleviates vaginal symptoms and reduces fungal burden in a murine vaginal candidiasis model. (A) Flowchart of mouse vaginal infection model. (B) Representative images of external vaginal signs (top) and gross reproductive anatomy (bottom) in mice from each group. (C) Representative images of C. albicans cellular morphology in vaginal lavage fluid from mice in each group. Abundant fungal cells and elongated hyphal forms were observed in both the infection model group and fluconazole monotherapy group. In contrast, after combined treatment with L. paragasseri , fungal burden was significantly reduced, and the morphological transition from yeast to hyphal forms was markedly inhibited. Scale bar, 10 μm. Images are representative of four independent experiments.

Article Snippet: Key experimental reagents included fluconazole (FLU, MedChemExpress), antibiotics (streptomycin, ampicillin, gentamicin, and kanamycin, Solarbio Technology Co., Ltd., Beijing, China), and 17β-estradiol benzoate (Yuanye Bio).

Techniques: Infection, Morphology, Yeast

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet: Fluconazole-resistant Candida auris thrives during zinc deficiency (A) In vitro evolution model of C. auris fluconazole resistance. (B) Growth percentage of C. auris under various concentrations of zinc chelator, normalized to the growth at 0 μM zinc chelator. (C) Growth of C. auris at 48 h under different concentrations of zinc chelator and normalized to 0 μM zinc chelator. The strains BJCA002 (clade III), CBS12766 (clade I), 2853 (clade I), and 1195 (clade I) are clinical isolates and are resistant to fluconazole. (D) The number of fungal cells ingested (phagocytic index) per 100 macrophages was manually counted at 3 h of co-incubation. MOI = 1, n = 3. (E–F) The survival of three strains of C. auris after infection of macrophages at an MOI = 1 for 3 h (E) and 12 h (F), evaluated by plating dilutions on YPD agar ( n = 3). Data are presented as the mean ± SD (B–F). Significance was determined using two-way ANOVA with Tukey's test (D–F). ns, not significant; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. Abbreviations: FLC, fluconazole; MOI, multiplicity of infection; WT, wild type; SD, standard deviation.

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: In Vitro, Incubation, Infection, Standard Deviation

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet: Modulation of zinc transporters contributes to the survival of fluconazole-resistant Candida auris in macrophages (A) C. auris was incubated for 4 h in YPD (Ctrl Zn) and YPD containing a zinc chelator (Low Zn). Zinquin was used to probe the zinc level, and representative images were shown. Scale bars = 5 μm. (B) C. auris was co-cultured with macrophages at MOI = 1 for 3 h, and the samples were stained with membrane-permeable fluorescent dyes Zinpyr green for labile zinc and DAPI for the nucleus. Images were examined using a confocal microscope. Scale bars represent 50 μm and 10 μm in the low- and high-magnification images, respectively. (C) Flow cytometric quantification of zinc level in infected macrophages after incubation for 3 h. MOI = 1, n = 3. (D–G) Expression of the Slc30a family (D) and Slc39a family (E) in macrophages following C. auris infection under low zinc conditions for 3 h, and expression of the Slc30a family (F) and Slc39a family (G) for 12 h. n = 3. The x -axis represents the various genes belonging to the Slc30a or Slc39a families. Data are presented as the mean ± SD (C–G). Significance was determined using two-way ANOVA with Tukey's test. ns, not significant; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. Abbreviations: MOI, multiplicity of infection; ND, not detected; SD, standard deviation.

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: Incubation, Cell Culture, Staining, Membrane, Confocal, Microscopy, Infection, Expressing, Standard Deviation

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet: Suppression of ZCF4 promotes fluconazole-resistant Candida auris survival during macrophage interaction (A) Transcriptomic dynamics of C. auris during infection under different zinc conditions. RNA sequencing analysis of C. auris gene expression (log 2 (fold change)) during infection, compared to the WT adapted to IMDM medium (time point 0), under varying zinc conditions. Lines represent log 2 (fold change) for individual genes, with numeric labels indicating significantly differentially expressed genes (DEGs) (|log 2 FC| > 1.5, adjusted P < 0.01; orange = upregulated, purple = downregulated). (B) Venn diagrams showing similarities and differences in the DEGs (|log 2 FC| > 1.5, adjusted P < 0.01) of C. auris in response to macrophages. (C) Clustering analysis of DEGs in C. auris under normal and low zinc conditions over a 3 h infection period. (D–E) mRNA expression of ZCF4 in C. auris was measured by RT-qPCR at 3 h (D) and 12 h (E) post infection. MOI = 1, n = 3. (F) Survival analysis of WT (CBS12766) or zcf4 mutant C. auris in macrophages at 3 h (left) and 12 h (right) post infection for MOI = 1. n = 6. (G) mRNA expression of ZRT2 (left) and ZRT3 (right) in C. auris was measured by RT-qPCR post infection. MOI = 1, n = 3. Data are presented as the mean ± SD (D–G). ns, not significant; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; by one-way ANOVA with Tukey's test (D–E) and two-way ANOVA with Holm–Šidák post hoc test (F–G). Abbreviations: MOI, multiplicity of infection; WT, wild type; SD, standard deviation.

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: Infection, RNA Sequencing, Analysis, Gene Expression, Expressing, Quantitative RT-PCR, Mutagenesis, Standard Deviation

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet: Fluconazole-resistant Candida auris impedes phagocytic clearance via inhibition of PI3K-AKT-MMP-9 pathway (A–B) mRNA of Mmp-9 in C. auris was measured by RT-qPCR at 3 h (A) and 12 h (B) post infection. MOI = 1, n = 3. (C) MMP-9 and MMP-2 gelatinase activity in the supernatant of C. auris and macrophage co-culture after 12 h of incubation. Mock, macrophages without C. auris infection. (D) Immunoblotting of MMP-9-related protein expression in C. auris -infected macrophages at 12 h. Macrophages were infected with C. auris , and cell lysates were prepared for immunoblotting with the indicated antibodies at 12 h. Phosphorylated proteins were normalized to total proteins, and target proteins were normalized to the loading control. Mock, macrophages without C. auris infection. (E) Survival analysis of C. auris in macrophages at MOI = 1 after 12 h with or without MMP-9 inhibitor under zinc limitation. n = 6. Data are presented as the mean ± SD. ns, not significant; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; by one-way ANOVA with Tukey's test (A–B) and two-way ANOVA with Šidák post hoc test (E). Abbreviations: act, activate; MMP, matrix metalloproteinase; MOI, multiplicity of infection; WT, wild type; SD, standard deviation.

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: Inhibition, Quantitative RT-PCR, Infection, Activity Assay, Co-culture Assay, Incubation, Western Blot, Expressing, Control, Analysis, Standard Deviation

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet: Zinc deficiency facilitates systemic candidiasis by fluconazole-resistant Candida auris (A) Survival rate of Drosophila (w 1118 ) after C. auris infection with 1000 CFUs (left) and 3000 CFUs (right). Data represent the mean of three independent experiments. (B) Survival rate of Drosophila (gut-specific dZnT63C RNAi) after C. auris infection with 1000 CFUs. Data represent the mean of three independent experiments. (C) Schematic representation of virulence and gut commensalism. The abundance of C. auris was determined as fungal load (log 10 (CFU/g)). Colonization of each strain was measured by counting the cell numbers present in fresh fecal pellets on 9, 12, 15, and 24 d post inoculation. (D) Quantitative fungal burden in the kidney ( n = 4 mice per group at each time point). Mice were pre-treated with different diets and infected with 5 × 10 7 CFUs of WT or R2, followed by euthanasia and fungal recovery at 6 and 10 d post infection. (E) Representative histopathology cross-sections of kidneys from C. auris -infected mice were stained with PAS (scale bar = 500 μm, up; scale bar = 10 μm, down) or HE (scale bar = 500 μm, up; scale bar = 10 μm, down) at 6 d post infection. (F) Representative immunofluorescence images of the kidney in WT or R2-strain-infected mice under a low-zinc diet at 6 d post infection. Blue, nucleus; green, C. auris ; red, macrophage (F4/80); magenta, MMP-9. Scale bars: 50 μm. (G) Quantitative fungal burden in the spleen, kidney, and brain ( n = 4 mice per group at each time point) after systematic infection with 5 × 10 7 CFUs of WT (CBS12766) or zcf4Δ mutant, followed by euthanasia and fungal recovery at 6 and 10 d post infection. (H) Representative histopathology cross-sections of kidneys from WT or zcf4Δ mutant-infected mice, stained with PAS (scale bar = 500 μm, up; scale bar = 10 μm, down) or HE (scale bar = 500 μm, up; scale bar = 10 μm, down) at 6 d post infection. (I) Production of the cytokines and chemokines in WT or zcf4Δ mutant C. auris -infected mice, analyzed by ELISA ( n = 4). Data are presented as the mean ± SD (A, B, D, G, I). ns, not significant; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; by the Log Rank test (A–B), two-way ANOVA with Tukey's test (D), and two-way ANOVA with Šidák post hoc test (G, I). Abbreviations: CFU, colony-forming unit; Abs, antibiotics; ELISA, enzyme-linked immunosorbent assay; WT, wild type; SD, standard deviation.

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: Drosophila, Infection, Counting, Histopathology, Staining, Immunofluorescence, Mutagenesis, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: hLife

Article Title: ZCF4 -dependent suppression of MMP-9 drives virulence in fluconazole-resistant Candida auris

doi: 10.1016/j.hlife.2026.04.004

Figure Lengend Snippet:

Article Snippet: To generate resistant mutants, the fluconazole-susceptible strain BJCA001 was subjected to a progressive drug-pressure regimen in RPMI 1640 medium (Gibco, California, USA) starting with a concentration of 32 μg/mL fluconazole (48 h per passage at 35 °C; TOPSCIENCE, Shanghai, China).

Techniques: