Double mutation of optn and p62 increases the susceptibility to Mm infection but combined overexpression has no additive effect. (A) Comparison of single and double optn/p62 mutants. Four different genotypes were infected with 200 CFU of Mm following the indicated experimental workflow and assessed for bacterial burden at 4 dpi (representative images shown). Quantification shows that optn/p62 double mutant embryos were hypersusceptible compared to the wild type and single mutants. (B) Comparison of separate and combined overexpression of optn/p62. One cell stage embryos were injected with 100 pg optn mRNA, 100 pg p62 mRNA, a combination of 50 pg optn mRNA and 50 pg of p62 mRNA, or buffer as a control. The subsequent workflow for infection and analysis of bacterial burden (representative images shown) was the same as in (A). Quantification shows that the combined overexpression of optn and p62 was less effective in reducing bacterial burden. Data are displayed as percentage difference to the control group set at 100% and are accumulated from two (A) or three (B) independent infection experiments (each data point representing an individual embryo), indicated with different colors. ns, non-significant, * p<0.05, ***p<0.001.
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