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. 2018 Aug 20;115(36):E8528–E8537. doi: 10.1073/pnas.1808302115

Fig. 7.

Fig. 7.

Impact of T6SS2 on crypt cocolonization. (A) Timeline for cocolonization experiment. Animals were exposed to one of two different inocula that compete the CFP-tagged ES114 strain against the YFP-tagged FQ-A001 wild-type (FQ1) or vasA_2 mutant (FQ1−). Animals were exposed to the inoculum for ∼22 h, and animals were fixed for imaging ∼44 h after the initial inoculum exposure. (B) Image of the light organ of an animal exposed to ES114 differentially expressing CFP or YFP: (Left) CFP+ YFP− crypts; (Center) CFP−YFP+ crypts; overlay of CFP, YFP, and bright-field images of light organ. (C) Map of crypt colonization counts and how images are scored. Crypt colonizations are labeled according to strain type: singly colonized, either YFP (green) or CFP (blue) or cocolonized, YFP+ CFP+ (checkered/pink). (D) Individual animal crypt colonization scores for indicated competitions between CFP-tagged ES114 (ES) and YFP-tagged FQ-A001 wild-type (FQ1) or vasA_2 mutant (FQ1−); each row represents an animal and each box represents a crypt. Blue boxes represent crypts that were scored as CFP+ YFP−. Green boxes represent crypts that were scored as CFP− YFP+. Pink boxes represent crypts that were scored as CFP+ YFP+. (E) Table summarizing animal and crypt cocolonization frequencies for each trial of each treatment. (F) High-magnification, confocal image of an individual crypt cocolonized with ES114 and FQ1−. (G) Number of crypts that were scored as CFP+ YFP−, CFP− YFP+, and CFP+ YFP+ for competitions between ES114 and FQ1 or between ES114 and the FQ1 vasA_2 mutant. Data from two different trials are shown for each competition. The proportion of CFP+ YFP+ crypts between different types of competitions were compared using a two-proportion z-test and an asterisk indicates P < 0.001. The inoculum levels and YFP/CFP ratios for trial 1 were 7,760 CFU/mL and 1.15, respectively, for ES vs. FQ1, and 16,160 CFU/mL and 1.04, respectively, for ES vs. FQ1−. The inoculum levels and YFP/CFP ratios for trial 2 were 6,020 CFU/mL and 0.87, respectively, for ES vs. FQ1, and 8,400 CFU/mL and 1.14 for ES vs. FQ1−.