Figure 3
From: RIP3 attenuates the pancreatic damage induced by deletion of ATG7
Autophagy-deficient mice showed increased activity of apoptosis and necroptosis. (a) Depletion of Atg7 increased pancreatic caspase-3 in 20-week-old Atg7Δpan mice. Caspase-3 quantitation and representative IF microphotographs of Atg7F/F (n=4) and Atg7Δpan (n=4) pancreatic tissue stained for DAPI (blue) and Caspase-3 (red) (anti-active caspase-3 ab2302, 1/50, scale bar=50 μm). (b) Reduced pancreatic Atg7 level increased the expression of caspase-8 in 12-week-old Atg7Δpan mice. Caspase-8 quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Caspase-8 (green) (anti-active caspase-8 Novusbio NB100-56116, 1/1000, scale bar=50 μm). (c) Increased the expression of caspase-9 in 12-week-old Atg7Δpan mice. Caspase-9 quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Caspase-9 (green) (anti-caspase-9 Novusbio NB100-56118, 1/1000, scale bar=50 μm). (d) Reduced pancreatic Atg7 level increased the expression of Bax in 12-week-old Atg7Δpan mice Bax quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Bax (green) (anti-Bax sc-526, 1/50, scale bar=50 μm). (e) Reduced pancreatic Atg7 level increased the expression of necroptotic protein Rip3 in 20-week-old Atg7Δpan mice. Rip3 quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Rip3 (green) (anti-Rip3 ab62344, 1/200, scale bar=50 μm). (f) Reduced pancreatic Atg7 level increased the expression of necroptotic protein Mlkl in 20-week-old Atg7Δpan mice. Mlkl quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Mlkl (green) (anti-Mlkl ab194699, 1/100, scale bar=50 μm). (g) Reduced pancreatic Atg7 level increased the expression of necroptotic protein Hmgb1 in 20-week-old Atg7Δpan mice. Hmgb1 quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7Δpan (n=5) pancreatic tissue stained for DAPI (blue) and Hmgb1 (red) (anti-Hmgb1 ab18256, 1/1000, scale bar=50 μm). Data are mean±S.E.M. for the numbers of animals as indicated in the graph, *P<0.05, **P<0.01
