Figure 1.

Keratin protein and mRNA levels in pancreata of keratin-overexpressing transgenic mice and in mice recovering from acute pancreatitis. mRNA (A) and protein (B) levels of normal nontransgenic mice (WT) and mice overexpressing WT mouse K8 (K8), human K18 (K18), and both K8 and K18 (K8/K18) are shown. A: mRNA was determined by real-time RT-PCR using previously described primers30 to the overexpressed keratins. Expression levels are shown as a ratio to WT mice for mK8, mK18, and to K18 mice for hK18 (n = 3 mice/genotype), and normalized to ribosomal L7 RNA. B: Equal amounts of total pancreas lysates from the mice analyzed in A were separated by SDS-PAGE and then immunoblotted with antibodies to the indicated keratins and actin. Three individual mice (nos. 1 to 3) were used for each genotype. C: Keratin protein levels in pancreata of nontransgenic (WT) mice and mice overexpressing mK8 (K8), hK18 (K18), and mK8 and hK18 (K8/K18) were compared to those in nontransgenic mice 2 days after acute pancreatitis injury. The nontransgenic mice were administered 7 hourly injections of caerulein (50 μg/kg of mouse weight) or of vehicle (saline) followed by isolation of the pancreas 48 hours after the first injection. Equal amounts of total pancreas lysates were separated by SDS-PAGE then immunoblotted with antibodies to the indicated proteins. Coomassie brilliant blue staining and the tubulin blot are included to show equal protein loading. Two age- and sex-matched individual mice (nos. 1 and 2) were used for each genotype.