Journal: Cell reports

Article Title: EGF-mediated suppression of cell extrusion during mucosal damage attenuates opportunistic fungal invasion

doi: 10.1016/j.celrep.2021.108896

Figure Lengend Snippet: (A) Heatmap of differentially expressed genesbetween homeostatic conditions (no MTZ) and induced epithelial cell loss by extrusion (MTZ-treated). q < 0.05, adjusted p value, Deseq2 and Benjamini-Hochberg method for FDR correction, three independent biological replicates, 15–30 larvae per replicate. (B) Gene Ontology (GO) analysis of gene categories enriched after epithelial cell loss. (C) Maximum intensity projections of fluorescent in situ hybridization for epigen in larvae with induced cell loss compared with homeostasis. Scale bar, 20 μm. (D–F) Larval survival after treatment with rhEPGN (D), rhEPGN and the EGFR inhibitor AG1478 (E), or rhEPGN and the MEK inhibitor U0126 (F) at different times during induction of damage and infection. Aggregated survival rates based on five (D, n = 72–74 per condition) or three independent experiments (E and F, n = 42–43 per condition), respectively, are plotted. Error bars represent SD; Mantel-Cox log rank test. (G) Quantification of the number of extruding cells with and without rhEPGN or rhEGF treatment during induced damage. Data are from three independent experiments, and error bars represent SD; ***p < 0.001; ordinary one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test. (H–K) Maximum intensity projections of confocal still images of a time-lapse sequence after induction of epithelial cell loss (H and I) and during treatment with rhEPGN (J) or rhEGF (K). Scale bar, 50 μm.

Article Snippet: Larvae were treated with either 30 nM rhEGF (Prospec Protein Specialists Product: CYT-217-b) or 250 nM rhEPGN (Peptides International Product: CYT-601) for a 24-h period prior to, during or immediately after 5 h MTZ (10 mM) treatment.

Techniques: In Situ Hybridization, Infection, Sequencing

Journal: Cell reports

Article Title: EGF-mediated suppression of cell extrusion during mucosal damage attenuates opportunistic fungal invasion

doi: 10.1016/j.celrep.2021.108896

Figure Lengend Snippet: (A–C) Maximum intensity projections of confocal images of larvae stained for the neutrophil marker mpx (green) during homeostasis (A), after mucosal damage (B), and mucosal damage and Rhizopus infection. Scale bar, 20 μm. (D) Quantification of the total number of neutrophils present in the epithelium (n = 108 number of animals). (E–G) Still images from time-lapse videos of neutrophil dynamics after cell loss-induced mucosal damage and after treatment with hrEPGN to suppress extrusion. Scale bar, 50 μm. (H) Quantification of the number of circulating neutrophils with and without rhEPGN or rhEGF treatment during induced damage. Data are from 38 animals from three independent experiments. Error bars represent SD. (I) Still images from time-lapse videos of a neutrophil migrating to a site of extrusion (denoted by asterisks). (J and K) Maximum intensity projection images of fluorescent in situ hybridization for mmp13a and il1b in larvae with induced cell loss compared with those treated with hrEPGN. Scale bar, 20 μm.

Article Snippet: Larvae were treated with either 30 nM rhEGF (Prospec Protein Specialists Product: CYT-217-b) or 250 nM rhEPGN (Peptides International Product: CYT-601) for a 24-h period prior to, during or immediately after 5 h MTZ (10 mM) treatment.

Techniques: Staining, Marker, Infection, In Situ Hybridization