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. 2017 Aug 18;85(9):e00347-17. doi: 10.1128/IAI.00347-17

FIG 4.

FIG 4

pdcA affects biofilm formation but not swimming motility or global c-di-GMP levels of C. difficile. (A) c-di-GMP in the cytoplasm of the 630 WT and pdcA::catP mutant C. difficile strains was quantified by UPLC-MS and normalized to the total cellular protein level. The mean values and standard deviations of six biologically independent samples are shown. (B) Swimming motility through BHIS–0.3% agar was measured at 24, 48, and 72 h. The mean values and standard deviations of six biologically independent samples are shown. (C) Biofilm formation assayed by crystal violet staining after 24 h of growth. The mean values and standard deviations of five biologically independent samples are shown. **, P < 0.01 by unpaired t test. (D, E) Complementation analysis of biofilm formation after 24 h of growth. pRT1099 is the vector control, pRT1214 encodes PdcA, and pRT1662 encodes a catalytically inactive form of PdcA. (D) Representative image of crystal violet-stained biofilms. (E) Quantification of biofilm biomass by crystal violet staining. The mean values and standard deviations of three biologically independent samples are shown. **, P < 0.05 by one-way analysis of variance and Dunnett's multiple-comparison test.