hela  (ATCC)

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: PAP1 expression under different growing conditions . Yeast-like cells were incubated under the following conditions at 37 °C: 1 h with either 1.0 μg mL −1 fibronection or thrombospondin 1; 1 h with a monolayer of HeLa cells; 1 h with 5 × 10 6 human PBMCs, or injected in the hemolymph of Galleria mellonella larvae and incubated for 24 h. Alternatively, biofilms were matured for 48 h at 37 °C. From these conditions, total RNA was extracted, cDNA synthesized with oligo(dT) primer (20 mer), and PAP1 expression quantified by RT-qPCR. Data were normalized using the expression of the gene encoding the ribosomal protein L6 and yeast-like cells growth in YPD at 37 °C as reference conditions (point zero on the Y axis). Results are means ± SD of three independent experiments performed in duplicate. The Dunnett's test and then the unpaired t -test were used for data analysis. * P < 0.05 when compared to the other growing conditions.

Article Snippet: In some experiments, HeLa cells (ATCC) were grown in monolayers in Eagle's Minimum Essential Medium (EMEM, Sigma-Aldrich) at 37 °C and 5 % CO 2 (v/v).

Techniques: Expressing, Incubation, Injection, Synthesized, Quantitative RT-PCR

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Adhesion to extracellular matrix components and HeLa cells of Sporothrix schenckii and Sporothrix brasiliensis PAP1 -silenced strains. The indicated extracellular matrix component was used to coat 96-well plates, then yeast-like cells were added, unbound cells were removed by extensive washing, and adherent cells were detected by ELISA with a primary anti-rHsp60 antibody. Control refers to wells coated with bovine serum albumin. Panel A contains data generated with S. schenckii, and WT is the 1099–18 ATCC MYA 4821 strain. Panel B shows results with S. brasiliensis cells, and the WT is the 5110 ATCC MYA 4823 strain. Alternatively, for both panels, 1 × 10 6 HeLa cells were placed per well, incubated 24 h at 37 °C and 5 % ( v /v) CO 2 , and used in the adhesion assays. Results are means ± SD of three biological replicates performed in duplicate. The Dunnett's test and then the unpaired t-test were used for data analysis. In both panels, * P < 0.05 when compared to WT or strains HSB1 and HSB2.

Article Snippet: In some experiments, HeLa cells (ATCC) were grown in monolayers in Eagle's Minimum Essential Medium (EMEM, Sigma-Aldrich) at 37 °C and 5 % CO 2 (v/v).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Generated, Incubation