Journal: Frontiers in Immunology
Article Title: The hypoxic microenvironment of Candida albicans biofilms shapes neutrophil responses
doi: 10.3389/fimmu.2025.1547559
Figure Lengend Snippet: Stabilization of HIF-1α in neutrophils following exposure to C albicans biofilms. (A) Representative Western blot result showing HIF-1α levels in neutrophils after 3 hours of co-culture with 48-hour C albicans biofilms formed by the ATCC 10231 and SC5314 strains. For HIF-1α analysis primary, rabbit antibodies anti-HIF-1α (1:1000, Cell Signaling) and secondary HRP-conjugated secondary antibodies (1:1000; R&D) were used. β-actin (mouse antibodies anti- β-actin, 1:10000, Cell Signaling) was used as a loading control. (B) Densitometric analysis of Western blot was performed in ImageJ software and presented as the relative levels of HIF-1α in neutrophils. The results were considered statistically significant for p < 0.05 (*p <0.05, ****p <0.0001). Data represent the results of five independent biological replicates (n=5). (C) Immunofluorescence staining of HIF-1α (red channel, Alexa Fluor 555) in neutrophils co-cultured with C albicans biofilms (ATCC 10231, SC5314, Δbgl2 , Δmnn9 and Δefg1/Δcph1 ). The image was also acquired using bright-field (BF) microscopy.
Article Snippet: For β-actin detection mouse anti-β-actin primary antibodies (1:10000, Cell Signaling Technology, Danvers, Massachusetts, USA; cat. #3700) were used.
Techniques: Western Blot, Co-Culture Assay, Control, Software, Immunofluorescence, Staining, Cell Culture, Microscopy