Figure 2
From: RIP3 attenuates the pancreatic damage induced by deletion of ATG7
Atg7 depletion induces chronic pancreatitis. (a) Representative H&E staining of pancreatic tissue demonstrated that loss of pancreatic Atg7 increased pancreatic tissue damage in Atg7Δpan mice (× 20 objective; scale bar=20 μm). (b) Histopathological evaluation of the pancreatic tissue included: For fibrosis: 0, no fibrosis; 1, mild fibrosis; 2, moderate fibrosis; 3, severe fibrosis. For vacuolization: 0, no vacuoles to 3, maximal vacuoles. For inflammation: 0, absent to 3, high infiltration. For Islets damage: 0, no islets damage to 3, severe islets damage, as described previously.2 (c) Loss of pancreatic Atg7 induces fibrosis. Atg7 quantitation and representative IF microphotographs of Atg7F/F (n=2) and Atg7Δpan (n=4) pancreatic tissue stained for DAPI (blue) and Collagen-1 (green) (anti-collagen, ab6308, 1/300, scale bar=50 μm). (d) Representative transmission electron microscopy image of pancreatic acinar cells from control and Atg7-depleted mice. Arrows indicate collagen fibers indicating tissue fibrosis after loss of pancreatic Atg7. (e) Loss of pancreatic Atg7 increased severity of fibrosis and ADM. Histopathology scores were age dependent plotted for fibrosis (continuous line) and ADM (broken line). (f) Induction of fibrosis was accompanied with an increased level of the pro-fibrotic factor TGFβ. TGFβ quantitation and representative IF/scattergram microphotographs of Atg7F/F (n=4) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and TGFβ (green) (anti-TGFβ ab66043, 1/100, scale bar=50 μm). (g) Depletion of pancreatic Atg7 increases tissue Trypsinogen/Trypsin expression. Trypsinogen/Trypsin quantitation and representative IF/scattergram microphotographs of Atg7F/F (n=4) and Atg7Δpan (n=4) pancreatic tissue stained for DAPI (blue) and Trypsinogen/Trypsin (green) (anti-Trypsinogen cs-67388, 1/50, scale bar=50 μm). (h) Loss of pancreatic Atg7 reduces survival. Survival rate was analyzed by using Kaplan–Meier analysis comparing survival in Atg7F/F (continuous line), Atg7Δpan (broken line) and Atg7Δpan-Rip3d/d (broken/continuous line). Data are mean±S.E.M. for the numbers of animals as indicated in the graph, *P<0.05, **P<0.01, ****P<0.0001
