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: 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: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibited ox-LDL-induced CD36 expression and Ca 2+ influx. (A) Histogram showing CD36 + macrophages in RAW264.7 macrophage after 100 µg/mL ox-LDL treatment were quantified by FCM. Statistical results of the proportion (B) and MFI (C) from (A) (n = 3). (D-E) The protein levels of CD36 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL and KAE(n = 3). (F) The fluorescence images of CD36 in different groups of RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL. (G) Statistical results of fluorescence images from (F) (n = 4). (H) Representative fluorescence images of Ca 2+ in different groups of RAW264.7 macrophage (I) Statistical results of fluorescence images from (H) (n = 4). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05 was compared with the ox-LDL group.

Article Snippet: Then, to prevent nonspecific binding, tissues 191 sections or cells were blocked with 4% bovine serum albumin for 1.5 h. Then, the tissues sections or cells were treated with primary antibodies, including IL-1β antibody (1:100, Wanlei), Piezo1 antibody (1:100, Proteintech), Nrf2 (1:100, CST), CD36 (1:100, CST) for 12 h at 4 °C.

Techniques: Expressing, Western Blot, Fluorescence, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibits macrophage inflammation and through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway. (A-B) The protein expression of CD36 by Western Blot in BMDMs after treatment (n = 3). (C) Representative fluorescence images of CD36 in different groups of BMDMs after treatment with 100 µg/mL ox-LDL. (D) Statistical results of fluorescence images from (C) (n = 4). (E-H) The protein expression of ERK, p-ERK, JNK, p-JNK, p38 and p-p38, by Western Blot in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). (I-J) The protein expression of p-p65, and p65 by Western Blot in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). (K-M) The protein expression of HO-1, and Nrf2 by in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). The results represent means ± SEM. *P < 0.05, **P < 0.01.

Article Snippet: Then, to prevent nonspecific binding, tissues 191 sections or cells were blocked with 4% bovine serum albumin for 1.5 h. Then, the tissues sections or cells were treated with primary antibodies, including IL-1β antibody (1:100, Wanlei), Piezo1 antibody (1:100, Proteintech), Nrf2 (1:100, CST), CD36 (1:100, CST) for 12 h at 4 °C.

Techniques: Expressing, Western Blot, Fluorescence

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: 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: 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