Journal: Nature Communications

Article Title: The machinery underlying malaria parasite virulence is conserved between rodent and human malaria parasites

doi: 10.1038/ncomms11659

Figure Lengend Snippet: ( a ) Survival of C57B/6 mice infected with mutant and wt parasites ( n =9 mice per group, P< 0.001, logrank test from Kaplan–Meier survival curve). ( b ) The course of parasitemia (mean±s.d.) in C57B/6 mice ( n =15 mice per group) infected with 10 5 mutant or wt parasites. Significant differences in parasitemia between wt and both mutants at day 3, 6, 7 ( P< 0.001) and day 5 ( P= 0.005) (two-tailed unpaired t -test). ( c ) Significant increase in number of mutant schizonts in the peripheral blood circulation compared with wt. Schizonts were quantified in Giemsa-stained smears of tail blood obtained 22 h after establishing synchronised infections (ratio of the number of mutant to wt schizonts, n =12 mice per group, P= 0.01; two-tailed unpaired t -test). ( d ) Representative distribution of sequestered schizonts in mice with synchronised infections 22 h post infection of mutant and wt parasites that express luciferase as shown by measuring luciferase activity (photons per s per cm 2 ; shown as radiance, y ; n =15 mice per group). ( e ) Quantification of parasite loads in isolated organs after perfusion 22 h after synchronised infections with mutant and wt parasites expressing luciferase. Parasite loads were determined by measuring luciferase activity (photons per s per cm 2 ) and expressed relative to wt ( n =15 mice per group). ( f ) Representative intravital images (spleen, fat) and ex vivo (lungs) confocal images of tissues from UBC-GFP mice 22 h after establishing synchronized infections of mutant and wt parasites that express mCherry. Insets in the images of fat tissue are enlargements of the yellow boxes. Arrowheads indicate parasites. Size bars, 10μm. ( g ) Spleens of mutant-infected mice are significantly larger (spleen-to-body weight ratio) than spleens of wt-infected mice ( n =9 mice per group, * P< 0.0001; unpaired Student's t - test; infections were done with 10 6 parasites per mouse). Images of representative spleens at day 4 after infection are shown to the right. ( h ) In vitro binding of schizonts of mutant and wt parasites to CD36-GFP-expressing CHO-745 cells (left) and to CD36 immobilized on plates (right). Binding is expressed as % of wt ( n =6, * P< 0.0001; two-tailed unpaired Student's t -test). ( i ) Quantitative FACS analysis of surface reactivity of RBCs infected with wt and gene-deletion mutant schizonts cultured ex vivo to mouse hyperimmune serum. Both deletion mutant schizonts exhibit significantly reduced levels of surface labelling with P. berghei sera compared to wt parasites. The geometric mean fluorescence intensity (MFI) relative to wt is shown (%). (*** P , 0.001, unpaired t -test).

Article Snippet: For in vitro measurement of the adhesion of mutant and wt P. berghei parasites to CD36-coated dishes recombinant mouse CD36/Fc chimera (purchased from R&D) was used.

Techniques: Infection, Mutagenesis, Two Tailed Test, Staining, Luciferase, Activity Assay, Isolation, Expressing, Ex Vivo, In Vitro, Binding Assay, Cell Culture, Fluorescence

Journal: Nature Communications

Article Title: The machinery underlying malaria parasite virulence is conserved between rodent and human malaria parasites

doi: 10.1038/ncomms11659

Figure Lengend Snippet: ( a ) The matched complemented mutants showed significantly increased binding to purified CD36 compared with the gene-deletion mutants (binding indicated as % of wt; * P< 0.0001, one-way ANOVA, Tukeys post hoc test.; n =3). ( b ) Survival of C57B/6 mice infected with matched complemented mutants was significantly increased in comparison with the mutants and the mismatched complemented mutants ( n =9 mice per group, P< 0.001, logrank test from Kaplan–Meier survival curve). ( c ) Significant decrease in number of schizonts in the peripheral blood circulation in matched complemented mutants compared with the gene-deletion mutants and to mismatched complemented mutants. Schizonts were quantified in Giemsa-stained smears of tail blood obtained 22 h after establishing synchronised infections ( n =9 mice per group; * P< 0.0001, one-way ANOVA, Tukeys post hoc test).

Article Snippet: For in vitro measurement of the adhesion of mutant and wt P. berghei parasites to CD36-coated dishes recombinant mouse CD36/Fc chimera (purchased from R&D) was used.

Techniques: Binding Assay, Purification, Infection, Staining

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