rabbit polyclonal anti cd36  (Novus Biologicals)

Journal: PLOS Pathogens

Article Title: Retinol binding protein 4 enhances cellular cholesterol uptake to facilitate influenza A virus infection

doi: 10.1371/journal.ppat.1013623

Figure Lengend Snippet: (A, B) Quantification of total cholesterol levels in multiple WT and RBP4-deficient cell types (A) and in serum from WT and RBP4-deficient mice (B). (C) Heatmap representation of RNA sequencing analysis showing differentially expressed genes related to cholesterol metabolism in WT and RBP4-deficient BMDMs. (D, E) qPCR analysis of CD36 mRNA expression in WT and RBP4-deficient BMDMs (D, left) and HEK293T cells (D, right), or in lung tissues from WT and RBP4-deficient mice (E). (F, G) Immunoblotting analysis of CD36 protein levels in multiple WT and RBP4-decficent cells as indicated (F) or in lung tissues from WT and RBP4-deficient mice (G). (H to J) Immunoblotting analysis of protein levels of CD36, total JNK, phosphorylated (p)-JNK, STAT1, and p-STAT1 as indicated in PK-15 or 3D4/21 cells pretreated with SP600125 (SP, 10 μM) or Resatorvid (TAK, 1 μM) for 2 hours, followed by treatment with rpRBP4 for 24 hours. (K) Immunoblotting analysis of protein expression of CD36, NP and M1 in PK-15 cells treated as described in (H), followed by infection with IAV WSN strain (MOI = 0.1) for 24 hours. (L) Immunoblotting analysis of CD36, p-STAT1, total STAT1 and RBP4 protein levels in HEK293T cells transfected with RBP4 expression plasmid or control empty vector, followed by treatment with Ruxolitinib (Ruxo, 5 μM) for 2 hours. (M) Immunoblotting analysis of CD36, p-STAT1, STAT1, STRA6 and RBP4 protein levels in HEK293T cells co-transfected with RBP4 expression plasmid and siRNA for STRA6 or negative control for 36 hours. (N) Immunoblotting analysis of CD36, p-JAK2, JAK2, p-STAT1, STAT1 and RBP4 protein levels in HEK293T cells transfected with RBP4 expression plasmid or control empty vector, followed by treatment with Ruxo (5 μM) for 2 hours or transfected with siRNA for STRA6 or negative control for 36 hours. Data are pooled from three independent experiments (A, B, D, E, mean ± SD) or representative of two independent experiments (F-N). * p < 0.05, ** p < 0.01 (Student’s t -tes t ).

Article Snippet: The primary antibodies used in this study were sourced from commercial suppliers as follows: Rabbit polyclonal antibodies against RBP4 (1 : 2,000, 11774–1-AP), CD36 (1 : 2,000, 18836–1-AP), STRA6 (1 : 3,000, 22001–1-AP), p65 (1 : 1,000, 10745–1-AP) and β-actin (1 : 20,000, 66009–1-lg) were purchased from Proteintech Group Inc. Rabbit polyclonal antibodies against CD36 (1 : 1,000, CY5796) and FLAG (1 : 10,000, AB0030) were purchased from Abways.

Techniques: RNA Sequencing, Expressing, Western Blot, Infection, Transfection, Plasmid Preparation, Control, Negative Control

Journal: PLOS Pathogens

Article Title: Retinol binding protein 4 enhances cellular cholesterol uptake to facilitate influenza A virus infection

doi: 10.1371/journal.ppat.1013623

Figure Lengend Snippet: (A to D) Confocal microscopy (A and B) or flow cytometry (C and D) analysis of cholesterol uptake by Dil-OxLDL in WT and RBP4-deficient HEK293T cells or BMDMs. Scale bars, 100 μm. Statistics of mean fluorescence intensity was calculated by Image J software (A and B, right) or Flow Jo software (C and D, right). (E, F) Flow cytometry analysis of cholesterol uptake by Dil-OxLDL in WT, RBP4-deficient and CD36 overexpressing RBP4-deficient HEK293T cells (E) and BMDMs (F). Mean fluorescence intensities were quantified with Flow Jo software. (G, H) Flow cytometry analysis of SA expression in HEK293T cells (G, upper) transfected with hCD36 plasmid or in stable CD36-overexpressing iBMDMs (H, upper). Cells were left untreated or incubated with cholesterol for 1 hour at 37°C, followed by staining with SNA or MAL. Mean fluorescence intensities were quantified by Flow Jo software (lower panels). (I) Confocal microscopy analysis of SA expression and distribution in WT and CD36-overexpressing iBMDMs. Cells were left untreated or incubated with cholesterol for 1 hour at 37°C, followed by staining with SNA or MAL (green). Cell membranes were stained with Dil dye (red), and nuclei were counterstained with DAPI (blue). Data are representative (A-F, left, G, H, upper and I) or pooled from at least three independent experiments (A-F, right, G and H, lower, mean ± SD). * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t -test).

Article Snippet: The primary antibodies used in this study were sourced from commercial suppliers as follows: Rabbit polyclonal antibodies against RBP4 (1 : 2,000, 11774–1-AP), CD36 (1 : 2,000, 18836–1-AP), STRA6 (1 : 3,000, 22001–1-AP), p65 (1 : 1,000, 10745–1-AP) and β-actin (1 : 20,000, 66009–1-lg) were purchased from Proteintech Group Inc. Rabbit polyclonal antibodies against CD36 (1 : 1,000, CY5796) and FLAG (1 : 10,000, AB0030) were purchased from Abways.

Techniques: Confocal Microscopy, Flow Cytometry, Fluorescence, Software, Expressing, Transfection, Plasmid Preparation, Incubation, Staining

Journal: PLOS Pathogens

Article Title: Retinol binding protein 4 enhances cellular cholesterol uptake to facilitate influenza A virus infection

doi: 10.1371/journal.ppat.1013623

Figure Lengend Snippet: (A, B) Flow cytometry analysis of α-2,3-linked SA or α-2,6-linked SA in WT, RBP4-deficient and CD36 overexpressing RBP4-deficient HEK293T cells, as wells as murine BMDMs transduced with CD36 retrovirus. Mean fluorescence intensities were quantified with FlowJo software. (C, D) qPCR analysis of vRNA levels of the NP gene or NP and M1 mRNA expression in HEK293T cells (C) or BMDMs (D) infected with IAV WSN strain (MOI = 5) during the viral attachment stage. (E) Flow cytometry analysis of α-2,3-linked SA or α-2,6-linked SA in WT and stable CD36-overexpressing iBMDMs cultured in serum-free medium. Cells were cultured with serum-free medium for 6 hours and subsequently stained with Lectins. Mean fluorescence intensities were quantified with FlowJo software. Data are pooled from three independent experiments (A-E, mean ± SD). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student’s t -test).

Article Snippet: The primary antibodies used in this study were sourced from commercial suppliers as follows: Rabbit polyclonal antibodies against RBP4 (1 : 2,000, 11774–1-AP), CD36 (1 : 2,000, 18836–1-AP), STRA6 (1 : 3,000, 22001–1-AP), p65 (1 : 1,000, 10745–1-AP) and β-actin (1 : 20,000, 66009–1-lg) were purchased from Proteintech Group Inc. Rabbit polyclonal antibodies against CD36 (1 : 1,000, CY5796) and FLAG (1 : 10,000, AB0030) were purchased from Abways.

Techniques: Flow Cytometry, Transduction, Fluorescence, Software, Expressing, Infection, Cell Culture, Staining

Journal: PLOS Pathogens

Article Title: Retinol binding protein 4 enhances cellular cholesterol uptake to facilitate influenza A virus infection

doi: 10.1371/journal.ppat.1013623

Figure Lengend Snippet: (A) Experimental scheme for lentiviral transduction and influenza virus infection in mice. WT and RBP4-deficient mice were intravenously injected with lentivirus (10 9 PFU in 200 μL per mouse) followed by infection with 10 4 PFU of IAV (WSN strain) 5 days later. Lungs were collected 3 days post-IAV infection. (B) qPCR analysis of CD36 mRNA levels in lung tissues from WT, RBP4-deficient, and CD36-overexpressing RBP4-deficient mice as treated in (A). (C) Body weight changes of indicated mouse genotypes at day 3 post-IAV infection. (D) Plaque assay analysis of virus titer in lung tissues from mice described in (A). (E, F) Immunofluorescence images (E, left) and quantitative analysis of NP protein intensity (E, right) or immunoblotting analysis of M1, NP and CD36 protein expression (F) in lung tissues from the mice described in (A). Mean fluorescence intensity was determined by Image J software (E, right). Scale bars, 100 μm. (G, H) Representative H&E-stained lung sections (G) and corresponding histopathological scores (H) from indicated groups. Each dot in (B-D, H) represents an individual mouse. (I) Proposed model of RBP4-mediated promotion of influenza virus infection. Created in BioRender. Huang, L. (2025) https://BioRender.com/rll2ll1 . Data are representative of two independent experiments (E-G). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student’s t -test).

Article Snippet: The primary antibodies used in this study were sourced from commercial suppliers as follows: Rabbit polyclonal antibodies against RBP4 (1 : 2,000, 11774–1-AP), CD36 (1 : 2,000, 18836–1-AP), STRA6 (1 : 3,000, 22001–1-AP), p65 (1 : 1,000, 10745–1-AP) and β-actin (1 : 20,000, 66009–1-lg) were purchased from Proteintech Group Inc. Rabbit polyclonal antibodies against CD36 (1 : 1,000, CY5796) and FLAG (1 : 10,000, AB0030) were purchased from Abways.

Techniques: Transduction, Virus, Infection, Injection, Plaque Assay, Immunofluorescence, Western Blot, Expressing, Fluorescence, Software, Staining

Journal: Communications Biology

Article Title: Fortilin deficiency induces anti-atherosclerotic phenotypes in macrophages and protects hypercholesterolemic mice against atherosclerosis

doi: 10.1038/s42003-025-08425-w

Figure Lengend Snippet: A.U. arbitrary unit, WT THP1 WT-fortilin cells, KO THP1 KO-fortilin cells, MTS 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H -tetrazolium, inner salt; BrdU bromodeoxyuridine, oxLDL oxidized low density lipoprotein, IB immunoblot, α-CD36 anti-CD36 antibody, ACs apoptotic cells, FarRed + cells labeled by CellTrace™ Far Red, CFSE + cells labeled by CellTrace™ CSFE. a DNA fragmentation assay of peritoneal macrophages (MΦ) from fortilin WT-MФ and fortilin KO-MФ mice when stimulated by vehicle or various concentrations of oxLDL (n = 5, 5; for each concentration of oxLDL). b DNA fragmentation assay of THP1 WT-fortilin (WT) and THP1 KO-fortilin (KO) cells with or without oxLDL stimulation (n = 3, 3). c MTS growth assay of THP1 WT-fortilin and THP1 KO-fortilin cells (n = 8, 8). d BrdU incorporation assay of induced peritoneal MФ from fortilin WT-MФ-HC and fortilin KO-MФ-HC mice (n = 6, 6). e FC quantification by Oil-Red-O staining of peritoneal MФ induced from fortilin WT-MФ-HC and fortilin KO-MФ-HC mice (n = 4, 4). f flow-cytometry-based FC formation assay using THP1 WT-fortilin and THP1 KO-fortilin cells (n = 3, 3). g JESS-based western blot analyses to assess the expression levels of CD36, an oxLDL receptor, with and without oxLDL stimulation, using THP1 WT-fortilin and THP1 KO-fortilin cells (n = 3, 3). h Efferocytosis assay. Workflow of the assay ①, representative flow cytometry plots ②, and efferocytosis indices of WT and KO cells ③ are shown. In ( e ), the scale bar equals 100 µm. Data are expressed as mean ± s.d., P values—determined by either two-sided Student’s t test ( a – f , h ) or one-way ANOVA with Tukey’s comparison ( g )—are shown.

Article Snippet: We used the following primary and secondary antibodies: Primary antibody: Rabbit anti-CD36 polyclonal antibody (Novus, St. Louis, MO, USA; Catalog #: NB400-145; 1:50 dilution) Secondary antibody: anti-rabbit secondary to horseradish peroxidase (HRP) mAb (ProteinSimple®, Catalog #: 042-206; 1:1 dilution).

Techniques: Western Blot, Labeling, DNA Fragmentation Assay, Concentration Assay, Growth Assay, BrdU Incorporation Assay, Staining, Flow Cytometry, Tube Formation Assay, Expressing, Comparison