Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: Enhanced hepatic VLDLR and CD36 levels in Sirt3 -deficient mice fed a HFD. mRNA abundance ( a ) and protein levels ( b ) of VLDLR in livers of WT and Sirt3 −/− mice fed either a standard chow or a HFD. CD36 ( c ), total and phospho-Nrf2 ( d ), Keap1 ( e ) and NQO1 ( f ) protein levels. g , ROS levels. h , PPARγ protein levels. Data are presented as the mean ± S.D. ( n = 6 per group). a, p < 0.05 vs. WT mice fed a standard chow. b, p < 0.05 vs. WT mice fed a HFD. c, p < 0.05 vs. Sirt3 −/− mice fed a standard chow

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: The increase in CD36 levels caused by lipids in Sirt3 -deficient hepatocytes is mediated by Nrf2. VLDLR mRNA abundance ( a ) and protein levels of VLDLR and NQO1, an Nrf-2-target gene, ( b ) were assessed in Huh-7 cells incubated with fatty acid free-BSA or BSA-palmitate (0.3 mM) and exposed to either vehicle or the Sirt3 inhibitor AAPBO (100 μM) for 16 h. a, p < 0.05 vs. CT. b, p < 0.05 vs. CT cells incubated with palmitate. c, p < 0.05 vs. CT cells treated with AAPBO. c , fatty acid uptake in Huh-7 cells incubated with fatty acid free-BSA or BSA-palmitate (0.3 mM) and exposed to either vehicle or the Sirt3 inhibitor AAPBO (100 μM) for 16 h was measured by the uptake of BODIPY-C16. a, p < 0.05 vs. CT. b, p < 0.05 vs. CT cells incubated with palmitate. c, p < 0.05 vs. CT cells treated with AAPBO. mRNA abundance ( d ) and protein levels of VLDLR ( e ) in Huh-7 cells transfected with control (CT) or SIRT3 siRNA and incubated in the presence or absence 0.3 mM palmitate (Pal) for 24 h. Protein levels of CD36 ( f ), NQO1 ( g ) and PPARγ ( h ) in Huh-7 cells transfected with control (CT) or SIRT3 siRNA and incubated in the presence or absence 0.3 mM palmitate (Pal) or the Nrf2 inhibitor ML385 (10 μM) for 24 h. a, p < 0.05 vs. CT siRNA cells. b, p < 0.05 vs. CT siRNA cells incubated with palmitate. c, p < 0.05 vs. SIRT3 siRNA cells. d, p < 0.05 vs. CT siRNA cells incubated with palmitate and ML385

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques: Incubation, Transfection, Control

Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: Potential new mechanisms by which Sirt3 deficiency promotes hepatic steatosis in mice fed a HFD. Exposure to a HFD reduces Sirt3 and contributes to triglyceride accumulation in the liver. However, hepatic lipid accumulation is attenuated by the activation of an adaptive mechanism involving an increase in nuclear HIF-1α and Lipin 1. Longer exposures to a HFD exacerbates Sirt3 decrease leading to higher uptake of lipids through an Nrf-2 mediated increase in CD36 and VLDLR. This results in a higher increase in fatty acid accumulation, which in turn reduces succinate levels, ultimately suppressing the adaptive increase in HIF-1α and Lipin 1. FAO: Fatty acid oxidation

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques: Activation Assay

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 3: MMP-9 regulates CD36 levels through proteolytic degradation. (A) Full length CD36

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 2: MMP-9 deletion increased CD36 and CILP levels at day 7 post-myocardial infarction

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 5: MMP-9 regulates phagocytosis through CD36. As shown by the in vivo stimulated

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques: In Vivo

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 8: Diagram depicting the roles of MMP-9 mediated CD36 degradation in LV remodeling

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Lipids in Health and Disease

Article Title: Accumulation of cholesterol, triglycerides and ceramides in hepatocellular carcinomas of diethylnitrosamine injected mice

doi: 10.1186/s12944-021-01567-w

Figure Lengend Snippet: Enzymes with a role in TG synthesis in the normal tissues (NT) and tumor tissues (TT) of mice injected with diethylnitrosamine. a Expression of ACC, pACC, FAS, and HSL. b Expression of DGAT1, DGAT2, pAMPK, AMPK and CD36. c Expression of LPL, PGC1alpha and LIPC. Cyclophilin A (CycA) was a further housekeeping protein analysed. d Ratio of pACC/ACC and pAMPK/AMPK in NT and TT. e Quantification of LPL protein. f Quantification of PGC1alpha protein. g Quantification of LIPC protein. h Quantification of proteins not changed in the TT. ( n = 6–7). * P < 0.05; *** P < 0.001

Article Snippet: Acetyl-CoA-carboxylase (ACC) (order number: MAB 6898), CD36 (order number: NB400-145), scavenger receptor BI (order number: NB400-104) and DGAT1 (order number: NB-110-4148755) antibodies were from Novus Biologicals (Wiesbaden-Nordenstadt, Germany).

Techniques: Injection, Expressing