v8 juice agar medium  (Thermo Fisher)

Journal: Applied and Environmental Microbiology

Article Title: A conserved C2H2 zinc finger protein, Rel1, links ribosome biogenesis to sexual development and antifungal susceptibility in a ubiquitous human fungal pathogen

doi: 10.1128/aem.01460-25

Figure Lengend Snippet: Rel1 played an important role in facilitating sexual reproduction. ( A ) Filamentation initiation assay of the WT and rel1 Δ strains under unisexual mating conditions. The mini-colonies of the indicated strains were photographed after 24 h of inoculation on V8 agar medium at 25°C in the dark. Scale bar, 100 µm. ( B ) The effect of REL1 deletion and complementation on hyphal development at the colony level during unisexual reproduction. Images were captured under a stereoscope after 5 days of incubation on V8 agar medium. Scale bars represent 200 µm for the upper panel and 1 mm for the lower panel, respectively. ( C ) Basidial maturation score assessment of the hyphae population from the WT, REL1 deletion, and complemental strains. Eighty basidia of each strain were calculated at the 7th day of inoculation on V8 medium. ***, P < 0.001; ns, not significant (two-tailed Student’s t -test). ( D ) Fluorescent observation of the Dmc1-mCherry expression signal in wild-type, rel1 disruption, and complementation strain background. Scale bar, 5 µm. ( E ) Quantitative assay of Dmc1-mCherry fluorescent intensity from the indicated strains after 7 days of incubation on V8 medium. Fluorescent signals of 65 basidia of each strain were calculated. ( F ) The impact of Rel1 on sporulation during unisexual reproduction. The images of hyphae tips with or without a spore chain were captured after 3 weeks of incubation on V8 medium. Scale bar, 10 µm.

Article Snippet: Unisexual/bisexual filamentation and sporulation assay were performed on V8 juice agar medium (0.5 g/L KH 2 PO 4 , 5% V8 juice [wt/vol], and 4% Bacto agar, pH 7.0) at a 25°C incubator in the dark.

Techniques: Incubation, Two Tailed Test, Expressing, Disruption

Journal: Applied and Environmental Microbiology

Article Title: A conserved C2H2 zinc finger protein, Rel1, links ribosome biogenesis to sexual development and antifungal susceptibility in a ubiquitous human fungal pathogen

doi: 10.1128/aem.01460-25

Figure Lengend Snippet: RNA-seq analysis revealed Rel1’s role in facilitating sexual reproduction. ( A ) Venn diagrams of the differentially expressed genes of the WT_V8/WT_YPD group and rel1 Δ_V8/WT_V8 group. (B, C) The top 20 enriched Gene Ontology (GO) terms among the upregulated genes in the WT_V8/WT_YPD group ( B ) and the downregulated genes in the rel1 Δ_V8/WT_V8 group ( C ) are presented. ( D ) Heatmap analysis of the genes involved in mating/cell fusion, filamentation, and meiosis/sporulation in the WT_V8/WT_YPD and rel1 Δ_V8/WT_V8 groups. Log2(fold change) of each gene derived from three biological replicates in the indicated groups is presented. ( E ) RT-qPCR assay of the key sexual development genes, ZNF2 , CFL1 , PUM1, and DMC1 , in WT, rel1 deletion, and complemented strains under mating-inducing conditions. Error bar indicates mean ± SD from three independent biological replicates. Statistical significance was determined by a two-tailed Student’s t -test (***, P < 0.001; **, P < 0.01; *, P < 0.05). ( F ) Cell-cell fusion assay of the indicated crosses on the double-drug selective plate after 14 h of incubation on V8 medium at 25°C in the dark. The mean relative cell fusion efficiency from three biological replicates of each cross is labeled at the bottom. ( G ) Quantification of the cell fusion efficiency of the indicated crosses from panel ( F ). Error bars represented mean ± SD from three biological replicates. Statistically significant differences are indicated by the asterisks (**, P < 0.01, two-tailed Student’s t -test). ND, not detected.

Article Snippet: Unisexual/bisexual filamentation and sporulation assay were performed on V8 juice agar medium (0.5 g/L KH 2 PO 4 , 5% V8 juice [wt/vol], and 4% Bacto agar, pH 7.0) at a 25°C incubator in the dark.

Techniques: RNA Sequencing, Derivative Assay, Quantitative RT-PCR, Two Tailed Test, Cell-Cell Fusion Assay, Incubation, Labeling