CD14 and ALPK1 Affect Expression of Tight Junction Components and Proinflammatory Mediators upon Bacterial Stimulation in a Colonic 3D Organoid Model

Figure 5

Expression of tight junction genes after E. coli Nissle stimulation. Wild-type (WT) and knockout organoids were stimulated with E. coli Nissle 1917 for 6 hours and analysed for changes in tight junction component gene and protein expression. Differences between knockout and WT organoids were investigated. Values were normalized to unstimulated controls within each genotype for each experiment and represent the means with 95% CI. Unstimulated: WT , Cd14^-/-, Il10^-/-, and Alpk1^-/-. Stimulated: WT , Cd14^-/-, Il10^-/-, and Alpk1^-/-. (a) Gene expression. RQ = relative quantification. (b) Detection of TJ proteins by Western blot analysis of lysates from organoids after 6 h of EcN stimulation. Densitometric quantification normalized to GAPDH was calculated as change from the unstimulated control. RD = relative density %. Stimulated versus unstimulated values were compared using unpaired -tests. Welch’s correction was used in case of unequal variances. One-way analysis of variance followed by Dunnett’s multiple comparisons was performed. Each knockout genotype was compared to WT. , , , and .

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