Figure 5. LS174T cells were treated with control BSA, 0.5 mM palmitate or 0.5 mM palmitate and 50 ng/mL of IL-22 for 24 hours.

mRNA levels of ER/oxidative stress markers (a) GRP78, (b) spliced XBP1 (c) NOS2, (d) goblet cell differentiation factor KLF4, (e) component of the glycocalyx cell surface MUC1, (f) major secretory product of goblet cells MUC2 were determined by qRT-PCR. Normalized to expression of β-Actin and expressed as a fold change of the mean of BSA controls. Statistics: n = 8 per group (2 individual experiments). Data presented as box plots with whiskers show median, Q1, Q3 and min/max. One-way ANOVA with Bonferroni post-test Con versus treatments. *P < 0.05 **P < 0.01 ***P < 0.001. (g) Identification of the MUC2 precursor and mature glycosylated proteins in cell lysates analyzed under reducing conditions by agarose gel electrophoresis and Western blotting with the human MUC2 antibody reactive with glycosylated MUC2 and 4F1 MUC2 nonglycosylated precursor antibody, densitometry shows n = 4–6 from 2 independent experiments, mean ± SEM. Unpaired student t test Con versus treatments; *P < 0.05 ***P < 0.001. Secreted MUC2 concentration shown as arbitrary units/mL were determined using an ELISA. n = 8. (i) Transepithelial electrical resistance was measured in control and treated cells after 24 h. qRT-PCR was used to determine the changes in the mRNA levels of ER stress marker (j) GRP78 and secreted gel-forming MUCIN-2 in human intestinal primary organoid cultures treated with control BSA, 0.5 mM palmitate or 0.5 mM palmitate and 50 ng/mL of IL-22 for 24 hours. n = 6. Data presented as box plots with whiskers show median, Q1, Q3 and min/max. One-way ANOVA with Bonferroni post-test Con versus treatments. *p < 0.05 **p < 0.01 ***p < 0.001.