Journal: Cell metabolism

Article Title: The MYC Oncogene Cooperates with Sterol Regulated Element-Binding Protein To Regulate Lipogenesis Essential for Neoplastic Growth

doi: 10.1016/j.cmet.2019.07.012

Figure Lengend Snippet: Upregulation of lipid metabolic genes by MYC. (A) Diagram of MYC’s regulation of the fatty acid synthesis pathway. (B) qPCR shows fatty acid synthesis mRNA expression in high MYC (MYC ON, N=3 per cell line) compared to 24 hours of MYC inactivation (MYC OFF, N=3 per cell line) in BCL (P493–6) and RCC (E28). Expression of lipogenesis proteins for BCL (P493–6) and RCC (E28) cells with MYC ON vs. MYC OFF, by Western blot analysis. Statistical significance by t-test, ***P<0.001. (C) RNA-seq analysis of lipid metabolic pathways in fold-change of RPKM comparing MYC ON/MYC OFF in BCL (P493–6).

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-SREBP1 (clone 2A4) [1:1000] home-made antibody N/A Rabbit monoclonal anti-MYC (Y69) [1:1000] Abcam CAS# ab32072 IgG from mouse serum [1:1000] Sigma CAS# I5006 IgG from rabbit serum [1:1000] Sigma CAS# I5381 MYC [1:1000] Abcam Ab32072 SREBP1 [1:1000] Novus Biologicals NB600–582 SCD1 [1:1000] Cell Signaling Tech CST 2794 FASN [1:500] Sigma-Aldrich F9554 ACLY [1:1000] Cell Signaling Tech CST 4332S ACACA [1:1000] Cell Signaling Tech CST 3661 Chemicals, Peptides, and Recombinant Proteins Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) Sigma-Aldrich CAS# C2920 Rotenone Sigma-Aldrich CAS# R8875 Seahorse XFe96 FluxPak Agilent Technologies CAS# 102416–100 XF Base Medium Seahorse Bioscience CAS# 102353–100 D-Glucose-1- 13 C Sigma 297046 L-Glutamine- 13 C 5 Sigma 605166 Critical Commercial Assays ChIP-IT® Express Chromatin Immunoprecipitation Kits Active Motif 53008 Qiagen RNA Extraction kit (RNEasy Plus Kit) Qiagen 74134 Deposited Data MYC Targeted Long Noncoding RNA DANCR Promotes Cancer in Part by Reducing p21 Levels.

Techniques: Expressing, Western Blot, RNA Sequencing Assay

Journal: Cell metabolism

Article Title: The MYC Oncogene Cooperates with Sterol Regulated Element-Binding Protein To Regulate Lipogenesis Essential for Neoplastic Growth

doi: 10.1016/j.cmet.2019.07.012

Figure Lengend Snippet: MYC regulates SREBP and together they regulate lipogenesis. (A) mRNA expression of fatty acid synthesis genes: Acly, Acaca, Fasn, and Scd1 upon siRNA knockdown of SREBP1 or scramble RNA in HCC(EC4) under MYC ON and OFF conditions (N=3 per condition). Statistical significance by two-way ANOVA using Tukey’s multiple comparisons post tests, ***P<0.001. Overall, ***P<0.001 for SREBP1 effect, ***P<0.001 for MYC effect and ***P<0.001 for interaction between the two. (B) SREBP1 ChIP-qPCR on BCL P493–6 line shows decreased binding of SREBP1 upon MYC inactivation (top panel). MYC ChIP-qPCR on HCC EC4 line shows no change of MYC binding upon SREBP1 knockdown (bottom panel). (C) Glucose and glutamine incorporation into lipids by NMR in MYC ON (upper panel) and OFF (bottom panel) in BCL(P493–6). For close-up look see Figure S3B. (D) Incorporation of glucose and glutamine carbons to de novo synthesized fatty acids by mass spectrometry based on the fold change of 2C13 peak in BCL(P493–6) as measured upon 24 hours of MYC inactivation (N=5). Fold change in the de novo fatty acid synthesis when MYC is activated or inactivated (left panel); and glucose and glutamine incorporation into fatty acids when MYC is activated (middle panel), or inactivated (right panel). Statistical significance by t-test, **P<0.01, ***P<0.001.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-SREBP1 (clone 2A4) [1:1000] home-made antibody N/A Rabbit monoclonal anti-MYC (Y69) [1:1000] Abcam CAS# ab32072 IgG from mouse serum [1:1000] Sigma CAS# I5006 IgG from rabbit serum [1:1000] Sigma CAS# I5381 MYC [1:1000] Abcam Ab32072 SREBP1 [1:1000] Novus Biologicals NB600–582 SCD1 [1:1000] Cell Signaling Tech CST 2794 FASN [1:500] Sigma-Aldrich F9554 ACLY [1:1000] Cell Signaling Tech CST 4332S ACACA [1:1000] Cell Signaling Tech CST 3661 Chemicals, Peptides, and Recombinant Proteins Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) Sigma-Aldrich CAS# C2920 Rotenone Sigma-Aldrich CAS# R8875 Seahorse XFe96 FluxPak Agilent Technologies CAS# 102416–100 XF Base Medium Seahorse Bioscience CAS# 102353–100 D-Glucose-1- 13 C Sigma 297046 L-Glutamine- 13 C 5 Sigma 605166 Critical Commercial Assays ChIP-IT® Express Chromatin Immunoprecipitation Kits Active Motif 53008 Qiagen RNA Extraction kit (RNEasy Plus Kit) Qiagen 74134 Deposited Data MYC Targeted Long Noncoding RNA DANCR Promotes Cancer in Part by Reducing p21 Levels.

Techniques: Expressing, Binding Assay, Synthesized, Mass Spectrometry

Journal: Cell metabolism

Article Title: The MYC Oncogene Cooperates with Sterol Regulated Element-Binding Protein To Regulate Lipogenesis Essential for Neoplastic Growth

doi: 10.1016/j.cmet.2019.07.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-SREBP1 (clone 2A4) [1:1000] home-made antibody N/A Rabbit monoclonal anti-MYC (Y69) [1:1000] Abcam CAS# ab32072 IgG from mouse serum [1:1000] Sigma CAS# I5006 IgG from rabbit serum [1:1000] Sigma CAS# I5381 MYC [1:1000] Abcam Ab32072 SREBP1 [1:1000] Novus Biologicals NB600–582 SCD1 [1:1000] Cell Signaling Tech CST 2794 FASN [1:500] Sigma-Aldrich F9554 ACLY [1:1000] Cell Signaling Tech CST 4332S ACACA [1:1000] Cell Signaling Tech CST 3661 Chemicals, Peptides, and Recombinant Proteins Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) Sigma-Aldrich CAS# C2920 Rotenone Sigma-Aldrich CAS# R8875 Seahorse XFe96 FluxPak Agilent Technologies CAS# 102416–100 XF Base Medium Seahorse Bioscience CAS# 102353–100 D-Glucose-1- 13 C Sigma 297046 L-Glutamine- 13 C 5 Sigma 605166 Critical Commercial Assays ChIP-IT® Express Chromatin Immunoprecipitation Kits Active Motif 53008 Qiagen RNA Extraction kit (RNEasy Plus Kit) Qiagen 74134 Deposited Data MYC Targeted Long Noncoding RNA DANCR Promotes Cancer in Part by Reducing p21 Levels.

Techniques: Recombinant, Chromatin Immunoprecipitation, RNA Extraction, Transgenic Assay, Software