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. 2020 Apr 13;6(4):e03717. doi: 10.1016/j.heliyon.2020.e03717

Figure 3.

Figure 3

Sporulation efficiency, morphological analysis, and transcript levels of spo0A in Clostridioides difficile strains (A) Sporulation efficiency (based on the reduction of non-germinated spores at 80 °C) was defined as the ratio of heat-resistant cells/total cells of each strain grown in the presence of 0.001% and 0.01% DCA compared to the cells grown without DCA (B) The cell membranes of C. difficile strains grown in the 70:30 broth were stained with FM4-64 and the double-stranded DNA was stained with Hoechst 33342. The samples were then observed under a fluorescence microscope (C) The stained cells were classified into four morphological stages, as shown in panel (B) and the number of each cell type was determined. One thousand cells were counted in each sample (D) The expression of spo0A in C. difficile strains grown for 24 h in 70:30 broth, with and without 0.01% DCA was analyzed using RT-qPCR. Values are relative to that in the absence of DCA (70:30 broth) after normalization to the transcript levels of the housekeeping 16S rRNA gene. Data are means ± standard deviation of triplicate experiments. ∗∗∗p < 0.001.