Journal: mBio
Article Title: Variations in candidalysin amino acid sequence influence toxicity and host responses
doi: 10.1128/mbio.03351-23
Figure Lengend Snippet: Candidalysin variants induce differential activation of MAPK signaling and cytokine secretion. Western blot analysis of TR146 epithelial cells treated with 15 µM of each candidalysin variant from C. albicans , C. dubliniensis, and C. tropicalis for ( A ) 30 min and ( B ) 2 h. Protein lysates (10 µg total protein) were probed with antibodies specific for MKK3, MKK6, and p-MKK3/6 ( A ), and p-EGFR, c-Fos, p-MKP1 ( B ). α-actin was used as a loading control. One representative blot presented from n = 3 biological repeats. Quantification of G-CSF, GM-CSF, IL-1α, IL-1β, and IL-6 secreted from TR146 epithelial cells treated with 15 µM of each candidalysin variant from ( C ) C. albicans , and ( D ) C. dubliniensis and C. tropicalis for 24 h. Data are the mean + SD of n = 3 biological repeats ( C. albicans candidalysin variant A vs variant B; no significant difference for all cytokines tested). Statistical analysis was applied relative to vehicle-treated cells using a one-way ANOVA with a post hoc Bonferroni multiple comparison test; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.
Article Snippet: Primary antibodies specific for MKK3 (#8535), MKK6 (#8550), p-MKK3/6 (MKK3-Ser189 and MKK6-Ser207; #12280), p-EGFR Y1068 (#3777S), c-Fos (#2250S), and p-MKP1 (#2857S) were purchased from Cell Signaling Technology. α-Actin (#MAB1501) was purchased from Millipore.
Techniques: Activation Assay, Western Blot, Variant Assay, Control, Comparison