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

Figure 3

Expression of tight junction after LPS stimulation. Organoids of different genotypes were stimulated with LPS for 6 hours. Change in tight junction component gene and protein expression was analysed and compared to wild type (WT). Values were normalized to unstimulated controls within each genotype in each experiment and represented the means with 95% CI and SEM. 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 LPS 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. , .

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