Pancreas-specific RelA/p65 truncation increases susceptibility of acini to inflammation-associated cell death following cerulein pancreatitis
J. Clin. Invest. Hana Algül, et al. 117:1490 doi:10.1172/JCI29882 [
Go to this article.]
Figure 6Impaired upregulation of murine PAP1 during AP in
relaΔ/Δ mice.
(
A and
B) Pancreata from
relaflox/flox and
relaΔ/Δ mice were removed at the indicated times. (
A) Total pancreatic RNA (8 μg;
n = 2) was labeled and hybridized to Affymetrix MOE430A GeneChips, and pancreas-specific genes were clustered hierarchically. (
B) Relative levels of PAP1 mRNA were determined by real-time PCR and expressed as mean ± SD (
n = 5). (
C) Pancreatic tissues were removed at the indicated times. Whole-tissue extracts were prepared and subjected to Western blot analysis using a newly generated antibody to murine PAP1. (
D) Chromatin immunoprecipitation experiments were performed with
relaΔ/Δ and
relaflox/flox pancreatic tissue at the indicated times after stimulation with cerulein using an antibody to p65. Precipitated DNA was analyzed by PCR using primers surrounding the positions of both κB sites in the respective promoters. PCR was also performed with 2.5% of input chromatin to ensure equal loading. (
E) Sections of snap-frozen pancreata and duodenum were prepared and analyzed for PAP1 expression in
relaflox/flox and
relaΔ/Δ mice in unstimulated pancreas and 12 and 24 hours after the first cerulein injection, respectively. Snap-frozen duodenum served as a positive control, because Paneth cells are known to express PAP1. Signals in the pancreas were localized to the apical regions of acini (arrows), typical for secretory proteins like PAP1.
