Figure 4 from Translational Activation of ATF4 through Mitochondrial Anaplerotic Metabolic Pathways Is Required for DLBCL Growth and Survival

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ATF4 protein level is decreased in vavP-Bcl2;Sirt3−/− mice and associated with lymphoma progression. A, Western blot results show the protein levels of ATF4, LC3, EIF2A, ACTB, and SIRT3 in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. The protein amounts were quantified with densitometry results. B, Summarized results of ATF4 protein level normalized to ACTB in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. The protein amounts were quantified with densitometry results from Western blots. C, Correlation between levels of autophagy (LC3II/LC3I) and ATF4 (ATF4/ACTB) in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. The data for correlation study were obtained with densitometry results from Western blots. D, Correlation between levels of phospho-EIF2A (p-EIF2A/EIF2A) and ATF4 (ATF4/ACTB) in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. The data for correlation study were obtained with densitometry results from Western blots. E, Summarized results of phospho-EIF2A level normalized to total EIF2A in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. The protein amounts were quantified with densitometry results from Western blots. F, Correlation between splenomegaly phenotype (spleen/body weight) and levels of ATF4 (ATF4/ACTB) in splenocytes from vavP-Bcl2;Sirt3+/+ and vavP-Bcl2;Sirt3−/− mice. ATF4 levels were quantified with densitometry results from Western blots. G, Western blot results show ATF4 levels from human DLBCL tumor samples or normal GC B cells from human tonsil. ACTB levels were used as loading control. H, GSEA shows the enrichment of ATF4 target genes in DLBCL tumors versus normal GC B cells. Gene expression data were from published microarray data (53). Error bars represent the mean ± SD of three or more replicates.

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