Macrophage-expressed Rac2 is sufficient to promote host survival against invasive A. fumigatus growth. rac2+/− and rac2+/−; mpeg1::mCherry-2A-rac2 adults were crossed to each other, and the resulting larvae were injected with YFP-expressing TBK1.1 (Af293) spores. (A) Survival was monitored for 7 dpi, and larvae were genotyped at the conclusion of the experiment for both the endogenous rac2 genotype and presence of the transgene. Average injection CFUs = 45. Data represent three pooled replicates. P values and hazard ratios were calculated by Cox proportional hazard analysis. (B–E) Larvae were live imaged via confocal microscopy at 1–5 dpi and genotyped at the conclusion of the experiment. Data represent 3 pooled replicates of 42 larvae per replicate. (B) The cumulative percentage of larvae experiencing germination was calculated. P values and hazard ratios calculated by Cox proportional hazard analysis. (C) Of larvae experiencing germination, the percentage that progress to invasive hyphae was calculated. P values calculated by Fisher’s exact test. (D) Fungal (YFP+) areas 0, 1, and 2 dpg were measured in larvae that experience germination at 3 dpi and displayed in a heatmap. (E) Example images showing germination in larvae at 3 dpi and subsequent invasive hyphae development and death (top) or control of fungal growth in larvae re-expressing Rac2 and mCherry in macrophages (bottom).
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