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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Cancer Discov. 2014 May 29;4(8):914–927. doi: 10.1158/2159-8290.CD-14-0363

Figure 6. Acute, systemic Atg7 deficiency compromises tumorigenesis.

Figure 6

A. Experimental design to induce lung tumors prior to conditional whole-body Atg7 deletion. Ubc-CreERT2/+;KrasG12D-frt/+;p53frt/frt;Atg7+/+ and Ubc-CreERT2/+;KrasG12D-frt/+;p53frt/frt;Atg7flox/flox mice were infected with Ad-FLPo at 6–8 weeks of age to activate RAS and delete p53, then these mice were treated with TAM at 12 weeks post-Ad-FLPo by intraperitoneal injection to create Atg7Δ/Δ mice. Tumor progression was analyzed at various times thereafter.

B. Representative micro-CT 3-dimensional reconstruction and quantification of lung volume showing healthy air space before TAM treatment and equivalent tumor burden at the time of TAM treatment of mice with the indicated genotypes. Kras+/+;p53+/+;Atg7+/+ mice were used as a control for normal lung airspace. Tumor histology and normal adjacent lung tissue (H&E images) were analyzed at 5 weeks post-TAM (last three panels). Arrows point to dead cells.

C. Quantification of wet lung weight (top) and tumor burden (bottom). Error bar represents SEM, p values are calculated using two-way ANOVA with Bonferroni post-test.

D. Representative western blotting for ATG7, p62, S6, P-S6 and LC3 of Atg7+/+ or AtgΔ/Δ tumor tissues. β-ACTIN serves a protein loading control.

E. Representative IHC for ATG7, p62, TOM20 and EM images of tumor tissue. Arrows in second panel point to p62 aggregates and arrows in the third panel point to TOM20 accumulation in tumors from ATG7-deficient mice. N: nucleus, M: mitochondria, Ly: Lysosome, AP: Autophagosome, L: Lipid.

F. Representative IHC images for active CASPASE-3 (left) with quantification (right). Error bar represents SEM, p values are calculated using two-way ANOVA with Bonferroni post-test.

G. Representative IHC images for KI67 (left) with quantification (right). Error bar represents SEM, p values are calculated using two-way ANOVA with Bonferroni post-test.

H. Representative IHC images for S6, P-S6, 4E-BP1, P-4E-BP1, ERK and P-ERK.

I. A model for destructive effect of autophagy on established tumors.