Journal: Gastroenterology

Article Title: A novel role for the dioxin receptor in fatty acid metabolism and hepatic steatosis.

doi: 10.1053/j.gastro.2010.03.033

Figure Lengend Snippet: Figure 3. Activation of AhR in transgenic mice induced hepatic expression of CD36 and uptake of fatty acids. (A) Real-time PCR analysis on the hepatic expression of CD36 mRNA in 2-month-old female mice. When applicable, mice were treated with DOX for 2 weeks before death. N 4 for each group. (B) Western blot analysis of the protein expression of CD36. Lanes represent individual mice. (C) Expression of CD36 and Cyp1a2 in WT and AhR-/- mice as determined by real-time PCR. (D) Real-time PCR analysis of the hepatic expression of Fatps, VLDLR, LDLR, SR-A, and SR-B. N 6 for each group. (E) FFA uptake in primary mouse hepatocytes was monitored by the uptake of BODIPY-C16. Top and bottom panels are fluorescence and phase-contrast images of the cells, respectively. DOX concentration is 1 g/mL. (F) Quantification of BODIPY-C16 uptake in panel E. AU, arbitrary unit. *P .05; **P .01, compared with control animals, or the comparisons are labeled.

Article Snippet: Priary antibodies used for Western blot analysis include poB100 (H-15) and ApoB48 (S-18) from Santa Cruz Santa Cruz, CA), CD36 (NB400-144) from Novus (Littleon, CO), AhR (SA-210) from Biomol (Plymouth Meeting, A), and -actin (A1978) from Sigma.

Techniques: Activation Assay, Transgenic Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Concentration Assay, Control, Labeling

Journal: Gastroenterology

Article Title: A novel role for the dioxin receptor in fatty acid metabolism and hepatic steatosis.

doi: 10.1053/j.gastro.2010.03.033

Figure Lengend Snippet: Figure 4. The mouse and human CD36 gene promoters are transcriptional targets of AhR. (A) Top: the sequences of mouse and human CD36 DREs and their mutant variants. Underlined are DREs and their mutants. Bottom: electrophoretic mobility shift assay results. (B) Chromatin immunoprecipitation assays to show the recruitment of AhR onto CD36 promoter. Huh-7 cells were treated with vehicle (Veh) or 10 nmol/L of TCDD (T) for 24 hours. CYP1A1/DRE and CYP7B1/non-DRE were included as a positive control and negative control, respectively. (C and D) Activation of the (C) mouse and (D) human CD36 promoter reporter genes by AhR in the presence of 3-methylcholanthrene (3-MC), or by CA-AhR without an exogenously added ligand. HepG2 cells were co-transfected with indicated reporters and receptors. Transfected cells were treated with vehicle (dimethyl sulfoxide) or 3-MC (2 mol/L) for 24 hours before luciferase assay. (E) Activation of mCD36 and hCD36 promoter reporter genes by endogenous AhR agonists Indigo (10 mol/L) and FICZ (200 nmol/L). *P .05; **P .01.

Article Snippet: Priary antibodies used for Western blot analysis include poB100 (H-15) and ApoB48 (S-18) from Santa Cruz Santa Cruz, CA), CD36 (NB400-144) from Novus (Littleon, CO), AhR (SA-210) from Biomol (Plymouth Meeting, A), and -actin (A1978) from Sigma.

Techniques: Mutagenesis, Electrophoretic Mobility Shift Assay, Chromatin Immunoprecipitation, Positive Control, Negative Control, Activation Assay, Transfection, Luciferase

Journal: Gastroenterology

Article Title: A novel role for the dioxin receptor in fatty acid metabolism and hepatic steatosis.

doi: 10.1053/j.gastro.2010.03.033

Figure Lengend Snippet: Figure 5. Loss of CD36 in mice inhibited the steatotic effect of an AhR agonist. (A and B) FFA uptake in primary mouse hepatocytes from WT and CD36/ mice treated with vehicle (Veh) or TCDD is monitored by the uptake of BODIPY-C16. (A) Top and bottom panels are fluorescence and phase-contrast images of the cells, respectively. (B) Quantification of BODIPY-C16 uptake in panel A. TCDD concentration is 10 nmol/L. (C and D) Two-month-old female mice were gavaged with a single dose of TCDD (30 g/kg). The mice were killed 7 days later and subjected to the measurements of (C) hepatic (n 4 for each group) and (D) plasma (n 5 for each group) triglyceride levels. Mice were fasted for 16 hours before killing. *P .05.

Article Snippet: Priary antibodies used for Western blot analysis include poB100 (H-15) and ApoB48 (S-18) from Santa Cruz Santa Cruz, CA), CD36 (NB400-144) from Novus (Littleon, CO), AhR (SA-210) from Biomol (Plymouth Meeting, A), and -actin (A1978) from Sigma.

Techniques: Concentration Assay, Clinical Proteomics

Journal: Gastroenterology

Article Title: A novel role for the dioxin receptor in fatty acid metabolism and hepatic steatosis.

doi: 10.1053/j.gastro.2010.03.033

Figure Lengend Snippet: Figure 7. Activation of AhR in transgenic mice inhibited hepatic fatty acid -oxidation, decreased white adipose tissue adiposity, and increased hepatic oxidative stress. (A) Suppression of PPAR and its target genes involved in fatty acid oxidation in 2-month-old female CA-AhR transgenic mice, as shown by real-time PCR analysis. N 6 for each group. (B) Inhibition of peroxisomal -oxidation in the liver extracts of transgenic mice. (C) Representative appearance of the omental fat, H&E staining of epididymal fat, and quantification of adipocyte size. (D) Omental fat tissue weight (white adipose tissue [WAT]) was measured as a percentage of total body weight (BW) in 2-month-old male WT and CD36/ mice gavaged with vehicle (Veh) or TCDD (30 g/kg) 7 days before being killed (n 5 for each group). (E) Abdomen adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) mRNA expression as measured by real-time PCR analysis (N 5 for each group). (F) Hepatic concentrations of malondialdehyde. Liver lipids were extracted from 5- to 6-week-old female mice and subjected to malondialdehyde measurement. When necessary, transgenic mice were treated with DOX for 2 weeks. N 3 for each group. *P .05; **P .01.

Article Snippet: Priary antibodies used for Western blot analysis include poB100 (H-15) and ApoB48 (S-18) from Santa Cruz Santa Cruz, CA), CD36 (NB400-144) from Novus (Littleon, CO), AhR (SA-210) from Biomol (Plymouth Meeting, A), and -actin (A1978) from Sigma.

Techniques: Activation Assay, Transgenic Assay, Real-time Polymerase Chain Reaction, Inhibition, Staining, Expressing

Journal: BioMed research international

Article Title: High Glucose Promotes CD36 Expression by Upregulating Peroxisome Proliferator-Activated Receptor γ Levels to Exacerbate Lipid Deposition in Renal Tubular Cells.

doi: 10.1155/2017/1414070

Figure Lengend Snippet: Figure 2: Effects of LY294002 on HG-induced CD36 expression. HK-2 cells were cultured with normal glucose (5.6 mM, NG), NG plus LY294002 (15 uM), high glucose (30 mM, HG), or HG plus LY294002 for 48 h. CD36 protein levels were determined by western blotting. Band intensities were normalized to 𝛽-actin band intensity using densitometry. The histogram represented the normalized intensities of proteins from three experiments. The data were represented as the means ± SD (a). CD36 mRNA levels were measured by RT-qPCR. 𝛽-Actin served as a reference gene. The results from three independent experiments were represented as means ± SD (b). ∗𝑃< 0.05 versus NG; #𝑃< 0.05 versus HG. Immunofluorescence staining of CD36 was shown in (c).

Article Snippet: Primary antibody against CD36 was purchased from Novus Biologicals, USA.

Techniques: Expressing, Cell Culture, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + orients naïve macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1β, IL12, TNFα, IL6, IFNγ, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Expressing, Membrane, Flow Cytometry, Co-Culture Assay

Journal: Cardiovascular research

Article Title: Beneficial effects of combinatorial micronutrition on body fat and atherosclerosis in mice.

doi: 10.1093/cvr/cvr146

Figure Lengend Snippet: Figure 2 Lipistase enhances muscle lipid utilization in LDLrKO mice. (A) Representative images of anti-CD36 immunostaining on paraffin-embedded sections of oxidative muscle (gastrocnemius) from mice after 10-month lipistase treatment vs. controls (bars ¼ 50 mm). (B) Oxygen consumption measured after the 10-month treatment (***P , 0.001). (C) Immunoblot quantification of the membrane-bound CD36 protein in muscle and mRNA expression of marker genes for fatty acid transport and oxidation measured in gastrocnemius 10 months after lipistase treatment (*P , 0.05).

Article Snippet: For CD36 quantification by immunoblotting, 4 mg and 6 mg of cytosolic and membrane extracts, respectively, were loaded and probed with 1/1000 rabbit polyclonal anti-CD36 antibody (Novus Biologicals, Inc. Littleton, CO, USA).

Techniques: Immunostaining, Western Blot, Membrane, Expressing, Marker

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: Current Zoology

Article Title: Context-dependent regulation of pectoralis myostatin and lipid transporters by temperature and photoperiod in dark-eyed juncos

doi: 10.1093/cz/zox020

Figure Lengend Snippet: Correlations among mass-independent metabolic rate residuals (from regressions of metabolic rate versus M b

Article Snippet: After transferring proteins onto membranes, we probed blots with antibodies against myostatin (goat polyclonal; R&D Systems, Minneapolis, MN, USA; 1:100 dilution), FAT/CD36 (rabbit polyclonal; Novus Biologicals, Littleton, CO, USA; 1:1,000 dilution), FABPpm (rabbit polyclonal, from Christopher G. Guglielmo; 1:10,000 dilution), FABP c (rabbit polyclonal, from Christopher G. Guglielmo; 1:8,000 dilution) , and GAPDH (chicken polyclonal; Millipore, Temecula, CA, USA; 1:8,000 dilution).

Techniques: Activity Assay, Expressing

Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 4. No protective effect of SS31 on GSH level and infarct size in CD36 KO mice. CD36 KO mice were subjected to 30 min MCAO. A, mice were treated with either saline (Veh) or SS31 (2 mg/kg body weight) immediately after reper- fusion, and GSH measurement was performed at 6-h post-ischemia. Values for GSHmeasurementareexpressedaspercentGSHdepletionintheipsilateralside compared with contralateral side. Error bars indicate S.D. (n 4 per group). B, mice were treated with either vehicle or SS31 (2 mg/kg body weight) immedi- ately after reperfusion, 6, 24, and 48 h. Infarct volumes were measured at 72-h post-ischemia. Error bars indicate S.D. (n 7 per group). No difference was observed between vehicle- and SS31-treated groups.

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Saline

Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 5. Effect of SS31 on ischemia-induced CD36 expression. C57BL/6 mice were subjected to 30 min MCAO and treated with saline (Veh) or SS31 (5 mg/kg body weight) immediately after reperfusion and again 6 h after ische- mia. A, for CD36 gene expression, total RNA was prepared from both hemi- spheres 24 h after ischemia, and CD36 mRNA level was determined. Error bars indicate S.D. (n 7). *, p 0.05 versus contralateral side; #, p 0.05 versus ipsilateral vehicle-treated group, one-way ANOVA with post-hoc Newman- Kuels test. B, correlation analysis of CD36 protein levels and infarct size. CD36 protein levels from SS31-treated MPM were expressed as arbitrary units. SS31 (5 mg/kg) was given at 0, 6, 24, and 48 h, and MPM were harvested 72 h after ischemia. Infarct volume was determined at 72 h after ischemia. CD36 protein levels were normalized against -actin levels. Note that CD36 protein level is positively correlated with infarct size (r 0.6390, p 0.0055).

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Expressing, Saline, Gene Expression

Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 6. Effect of SS31 on oxLDL-induced CD36 expression in MPM. Thioglycolate-elicited MPM were cultured and incubated with saline, oxLDL, andSS31peptidessinglyorincombinationasindicated.A,CD36mRNAlevels were determined 48 h after treatments. Error bars indicate S.D. (n 6 per group) *, p 0.05 versus V; #, p 0.05 versus oxL. B, CD36 protein levels were determined 48 h after treatments by Western blot analysis. Experiments were performed four times (n 4 per group). For each set of experiment, CD36 protein band densities were normalized against -actin. A vehicle-treated blot was used as a reference standard (100%), and CD36 band intensity was calculated based on the density of vehicle-treated blot. H, heart (positive control); KO, CD36 KO brain (negative control); Veh, saline; oxL, 25 g/ml oxLDL; SH, 106 M SS31; SL, 108 M SS31; #, p 0.05 versus oxL, one-way ANOVA with post-hoc Newman-Kuels test.

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Expressing, Cell Culture, Incubation, Saline, Western Blot, Positive Control, Negative Control