Fig. 1.

Intestinal epithelial-specific K8 deletion induces local keratin loss in intestinal epithelia. A Lysates of crudely isolated colon epithelium from K8^flox/flox (lane 1–3), K8^flox/flox; Villin-Cre (lane 4–6) and K8^flox/–; Villin-Cre (lane 7–9) mice (n = 3) were immunoblotted for K8, K7, K18, K19 and K20. B Total colon lysates from untreated K8^flox/flox; Villin-CreER^t2 (lane 1–3), and tamoxifen-treated (25 days after first injection) K8^flox/flox; Villin-CreER^t2 (lane 4–6) and K8^flox/flox (lane 7–8) mice (n = 3) were immunoblotted for K8, K7, K18, K19 and K20. Hsc70 was used as a loading control for both A and B. C, D The immunoblots in A and B were quantified and normalized to Hsc70. The results represent the mean (n = 3) protein quantity ± SD with significant differences shown between K8^flox/flox and K8^flox/flox; Villin-Cre mice (C), and between untreated and tamoxifen-treated K8^flox/flox; Villin-CreER^t2 mice (D), with individual values shown as dots. E, F The mRNA levels of Krt8, Krt7 (E and F) and Krt18, Krt19, Krt20 and Krt23 (E) of intestine-specific K8 knockout mice total colon lysates were analyzed by qRT-PCR. The results were normalized to both Actb and 18S ribosomal RNA expression and boxes (E) extend from 25 to 75th percentiles and line represents median expression value and whiskers represent min and max values with individual mice values shown as dots (n = 6), while (F) shows the average (n = 3) fold change ± SD and individual values shown as dots. G, H Ileum and I, J liver total lysates from intestine-specific K8 knockout mice were immunoblotted for K8 and Hsc70 was used as a loading control. The statistical significance was determined after one-way ANOVA, followed by post hoc Tukey multiple comparison test, expect in F by student’s T test, and shown as *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001