Fig. 5. Bap1 and RbmC are required for colonizing enteroid monolayers.

a Schematic for the growth of in vitro enteroid monolayers and subsequent colonization by V. cholerae cells. Created with Biorender.com. b Side (Top) and tilted view (Bottom) of a representative monolayer stained with DAPI and an F-actin probe conjugated to fluorescent Alexa Fluor^TM 647 dye (magenta) and colonized by V. cholerae biofilms from a rugose (Rg) strain constitutively expressing mNeonGreen. The total size of the view is 439$\times$439$\times$13 µm. c Representative results of a monolayer colonized by V. cholerae biofilms, before (Top) and after (Bottom) washing, showing defective colonization of the ΔrbmCΔbap1 mutant. Red dotted line demarks the boundary of the monolayer. d Quantification of monolayer colonization of different biofilm mutants (mean ± SD, n = 4 biologically independent samples, unpaired, two-tailed t-test with Welch’s correction. *p < 0.05 (Rg v.s. Δ rbmC: p = 0.0193; Δbap1 v.s. Δbap1 rbmC_ΔC1C2: p = 0.0114), **p < 0.01 (Δbap1 v.s. Δbap1 rbmC_ΔΜ1Μ2ː p = 0.0042), ***p < 0.001 (Rg v.s. Δbap1ΔrbmC: p = 0.0004), ns = not significant (Rg v.s. ΔvpsL: p = 0.4730; Rg v.s. Δbap1: p = 0.1107; ΔrbmC v.s. ΔrbmC bap1_Δ57aa: p = 0.1430).