Fluorescent-activated cell sorting analysis (FACS) indicates that the spores appear haploid when compared to the reference strains of C. neoformans JEC21 (haploid) and XL1549 (diploid) (FACS results shown in supporting File S1).
In NC4, we found a decrease in the number of spores, a delay in germination of the spores and delayed start of cell growth of the spores formed in space when compared to the ground control.
Infant botulism occurs because infants produce reduced amounts of bile acids, which inhibit the germination of C. botulinum spores, within the gastrointestinal tract when compared to the bile acids formed in an adult, and infant intestinal flora is immature.
Wild-type spores prepared at room temperature showed no difference in germination ability when compared to wild-type spores prepared at 37°C, suggesting that this was a reasonable method of complementation (data not shown).
Coat-depleted spores exhibited the expected enhanced sensitivity to lysozyme (ranging from 40 70%), when compared to non-stripped spores (about 10%) (data not shown).
Generally, spores are of different sizes, and spore viability is significantly reduced (1.81±1.46%) even when compared to rec12Δ mutants (20.99±5.17%) that fail to induce prDSBs (supplementary material Table S3).
There is no significant difference between the frequency of sister and non-sister spores in the sgs1-top3-id/sgs1Δ cross (ADA 12) when compared to wild type (Table 4).
After consecutive transfers of the mycelium from the single spore isolate, this marked effect was reduced and only a slight growth reduction was detectable when compared to the wild type (Figure 6).
Analysis of spore internalization showed that expression of Δp85α significantly decreased the spore internalization frequency (by ∼45%) in both A549 (Fig. 2, E, **, p<0.01) and HeLa cells (Fig. S1, D, ***, p<0.001) when compared to cells transfected with the vector control.
