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: 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: 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: 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: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: Visualization of molecular docking results using interaction diagrams. ( A ) Molecular docking of CAT and ERK1. ( B ) Molecular docking of CAT and MMP2. ( C ) Molecular docking of CAT and NOX4. ( D ) Molecular docking of CAT and RAGE. ( E ) Molecular docking of CAT and S100A8.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: CAT downregulated the S100A8-RAGE-NOX4 signaling pathway in HFpEF. ( A ) Representative WB bands of S100A8, RAGE, NOX4, ERK, p-ERK, P65, p-P65, and MMP2. ( B – G ) Quantification of protein expression levels analyzed using ImageJ software (n=3). ( H ) Quantitative analysis of IL-1β expression (n=3). ( I ) Quantitative analysis of IL-6 expression (n=3). ( J ) Quantitative analysis of TNF-α expression (n=3). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Software

Journal: The journal of physical chemistry. B

Article Title: Myelin-associated MAL and PLP are unusual among multi-pass transmembrane proteins in preferring ordered membrane domains

doi: 10.1021/acs.jpcb.0c03028

Figure Lengend Snippet: Shown are the raft affinity of multi-pass TMPs with varying number of TMHs. Raft affinity is shown in log scale, with 0 representing equal partitioning between the phases and positive and negative values representing enrichment in or depletion from the raft phase, respectively. Included in each panel is the value for LAT (a raft-preferring single pass TMD, black) and LDLR (a non-raft TMD, white). (A) 2-TM proteins. Three homologs of MOG from different species (h:human; m:mouse; r:rat) and both isoforms (alpha and beta) of the human protein are excluded from rafts, as is the unrelated CD36. (B) Two 4-TM proteins are highly raft-preferring, namely MAL and PLP. In contrast, several other members of the MARVEL family (BENE, MAL2, occludin, and synaptophysin) and other tetraspanin family proteins (CD9, CD81, CD82), as well as the neuronal membrane glycoprotein (GPM6a) are almost completely excluded. Two analogs of MAL (c:canine: h:human) were tested and both had significantly greater raft affinity than LAT. (C) All tested 6-TM and 7-TM proteins are raft excluded. Several proteins from the TRP family (TRPA1, TRPM8, TRPV1), Adenylyl Cyclase 9 (AC9), and G Protein-Coupled Receptors (DRD1, β2AR) were not significantly different from LDLR. (D) All tested proteins with 10 or more TMDs - PM scramblase TMEM16F and several transporters (NKCC1, KCNQ1 and GAT1) - were raft excluded. Data points represent the means of individual experiments of >10 vesicles each.

Article Snippet: Several multi-pass protein constructs were purchased from Addgene: mCherry-CD36-C10 (CD36, 55011), mCherry-Occludin-N-10 (Occludin, 55112), mCherry-CD9–10 (CD9, 55013), mCherry-CD81–10 (CD81, 55012), YFP-CD82 (CD82, 1819), pFCK(1.3)GFP-SynaptophysW (Synaptophysin, 27235), pEGFP-N-Drd1 (DRD1, 104358), Kv7.1/pcDNA3.1 (KCNQ1, 111452), pcDNA3.1(+)mGAT1–0-GFP (GAT1, 41662).

Techniques:

Journal: bioRxiv

Article Title: An evolutionary transcriptomics approach links CD36 to membrane remodeling in replicative senescence

doi: 10.1101/294512

Figure Lengend Snippet: Characterization of genes associated with replicative senescence. (A) The cumulative fraction plot of pLI (the probability of being loss-of-function intolerant) value of genes upregulated in old cells (YO, blue) and other protein-coding genes (gray). This measurement is obtained from ExAC database . (B) The cumulative fraction plot of b-value (the index of background selection between primate species) of genes in each category (Mc ). The color scheme is same as the previous panel. (C) X-axis shows the tolerance of genes in healthy human genomes (-log p- value of pLI), y-axis shows the log 2 fold change between young and old in BJ cells and the size of the bubble indicates the log 2 fold change between young and old in MRC-5 cells. The strength of the color shows the -log p-value of pLI. CD36 showed upregulation in both old BJ and MRC-5 cells and tolerance to loss-of-function mutations in healthy human genomes.

Article Snippet: FusionRed-CD36-C10 plasmid (#56104) was obtained from Addgene (deposited by the Davidson lab).

Techniques: Selection

Journal: bioRxiv

Article Title: An evolutionary transcriptomics approach links CD36 to membrane remodeling in replicative senescence

doi: 10.1101/294512

Figure Lengend Snippet: CD36 overexpression results in senescence-like phenotype. (A) Representative images of control and CD36 -overexpressing young MRC-5 cells. CD36 o verexpression was visualized via the FusionRed tag (scale bar = 10µm). (B-C) Measurement of β-galactosidase activity in control and CD36 overexpressing MRC-5 cells. A significant increase of SA-β-galactosidase positive cells was observed during CD36 overexpression for which representative images are shown (scale bar =100µm for control and center CD36 images, and 20µm for furthest to the right CD36 image). Data shown as means ± standard deviation, *** p < 0.001. (D) Representative western blot showing p16 levels in control and CD36 overexpressing young MRC-5 cells.α-tubulin was used as a loading control. We observe a modest increase in p16 levels. (E) Measurement of β-galactosidase activity in young MRC-5s that were treated with conditioned media. “Control” represents cells in standard growth medium, “Control_M” represents cells in medium from control wells during transfection, “+CD36_M1” represents cells in medium from CD36 overexpressing cells 48h after transfection, and “+CD36_M2” represents cells in medium from CD36 overexpressing cells 96h after transfection. An increase in SA-β-galactosidase positive cells was observed.

Article Snippet: FusionRed-CD36-C10 plasmid (#56104) was obtained from Addgene (deposited by the Davidson lab).

Techniques: Over Expression, Activity Assay, Standard Deviation, Western Blot, Transfection