Journal: Archives of biochemistry and biophysics

Article Title: ALL THREE HUMAN SCAVENGER RECEPTOR CLASS B PROTEINS CAN BIND AND TRANSPORT ALL THREE MACULAR XANTHOPHYLL CAROTENOIDS

doi: 10.1016/j.abb.2017.09.013

Figure Lengend Snippet: Immunohistochemistry showing expression of SR-B1 and CD36 in macaque eye sections. SR-B1 expression (red) was detected throughout the RPE and retinal layers. CD36 expression (red) was limited to the RPE layer. Both CD36 and SR-B1 antibodies were rabbit polyclonal. “No primary antibody” control sections were incubated with the secondary antibody alone. Nuclei are stained using DAPI in blue.

Article Snippet: The antibodies and their dilutions were as follows - 1:100 dilution of rabbit monoclonal SRB1 (ab52629-Abcam), 1:500 dilution of goat polyclonal anti-SRB2 (af1966-R&D Systems), and 1:100 dilution of rabbit monoclonal anti-CD36 (14347, Cell Signaling Technology).

Techniques: Immunohistochemistry, Expressing, Incubation, Staining

Journal: Archives of biochemistry and biophysics

Article Title: ALL THREE HUMAN SCAVENGER RECEPTOR CLASS B PROTEINS CAN BIND AND TRANSPORT ALL THREE MACULAR XANTHOPHYLL CAROTENOIDS

doi: 10.1016/j.abb.2017.09.013

Figure Lengend Snippet: SR-B2 staining in macaque eye sections. Protein expression was strong in the RPE layer and in the ganglion cell layer. Weak expression was detected throughout the retina. SR-B2 expression (red) was determined using a goat polyclonal antibody. “No primary antibody” control sections were incubated with only the secondary antibody. Nuclei are stained using DAPI in blue.

Article Snippet: The antibodies and their dilutions were as follows - 1:100 dilution of rabbit monoclonal SRB1 (ab52629-Abcam), 1:500 dilution of goat polyclonal anti-SRB2 (af1966-R&D Systems), and 1:100 dilution of rabbit monoclonal anti-CD36 (14347, Cell Signaling Technology).

Techniques: Staining, Expressing, Incubation

Journal: Asian Journal of Pharmaceutical Sciences

Article Title: Targeted delivery of glucocerebrosidase to lysosomes: The LYSOTAC (LYSOsome-TArgeting Chimera) technology

doi: 10.1016/j.ajps.2026.101149

Figure Lengend Snippet: LYSOTAC partially recovers GCase activity in LIMP-2 deficient HEK293 cells. (A) Scheme of generating SCARB2 -/- clones (Created in BioRender. Park, J. (2026) https://BioRender.com/6qf7ykt ); (B) WB of HEK293 cell clones transfected with gRNA vector targeting SCARB2 ; Relative quantification of (C) LIMP-2 and (D) GCase compared to control HEK293 cells in (B) ( n = 5); (E) GCase activity of HEK293 cell clones transfected with gRNA vector targeting SCARB2 ( n = 5); GCase activity of LIMP-2 deficient HEK293 cells (clone #2) exposed to (F) ATB2057 or (G) ATB2058 for 7 d was compared with control HEK293 cells ( n = 5); (H, I) Increased GCase activity of LIMP-2 deficient HEK293 cells in (F) and (G) is shown as a percentage change ( n = 5); (J) GCase activity of LIMP-2 deficient HEK293 cells treated with ATB2057 (1 µM), ATB1021 (1 µM), JZ-4109 (1 µM), and ambroxol (1 or 50 µM) for 7 d. Differences between groups were evaluated by one-way ANOVA (C, D, E, H, I, and J) or two-way ANOVA (F and G). The data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: LIMP-2-deficient HEK293 cells were produced using a LIMPII (SCARB2) Human Gene Knockout Kit (CRISPR) (# KN402162 ; Origene, Rockville, MD, USA) according to the manufacturer’s instructions.

Techniques: Activity Assay, Clone Assay, Transfection, Plasmid Preparation, Quantitative Proteomics, Control

Journal: BioMed Research International

Article Title: Delivery of Human EV71 Receptors by Adeno-Associated Virus Increases EV71 Infection-Induced Local Inflammation in Adult Mice

doi: 10.1155/2014/878139

Figure Lengend Snippet: Expression of recombinant AAV-hSCARB2 and AAV-hPSGL1 in cell lines increased susceptibility to EV71 infection. (a) Green fluorescence protein (GFP), human PSGL1 (hPSGL1), and human SCARB2 (hSCARB2) were cloned into the pEMBL plasmid with a CB promoter (pEMBL-CB). (b) L929 cells were transfected with pEMBL-CB-hPSGL1 and the expression of hPSGL1 was detected by surface stained with anti-hPSGL1 antibody and FITC-conjugate anti-mouse IgG and then analyzed by flow cytometry. (c) L929 cells were transfected with pEMBL-CB-hSCARB2 and the expression of hSCARB2 was detected by surface stained with anti-human SCARB2 and FITC-conjugate anti-mouse IgG and then analyzed by flow cytometry. (d) L929 cells transfected with the vector pEMBL-CB (Mock), pEMBL-CB-hSCARB2 (hSCARB2), and pEMBL-CB-hPSGL1 (hPSGL1) were infected with live EV71 (MOI = 1.0). Forty-eight hours after infection, protein extracted from cells was analyzed by Western blot using anti-EV71 VP1 and anti-actin antibodies. (e) Identical L929 cell treatment as described in (d) was observed for the cytopathic effect (CPE) by microscopy. Cells with CPE are labeled with a black arrow.

Article Snippet: Plasmid pCMV-hSCARB2 containing the human SCARB2 gene was purchased from OriGene Technologies, Inc. (Rockville, MD).

Techniques: Expressing, Recombinant, Infection, Fluorescence, Clone Assay, Plasmid Preparation, Transfection, Staining, Flow Cytometry, Western Blot, Microscopy, Labeling

Journal: Journal of Lipid Research

Article Title: Action myoclonus-renal failure syndrome: diagnostic applications of activity-based probes and lipid analysis

doi: 10.1194/jlr.m043802

Figure Lengend Snippet: Fig. 1. Visualization of GBA with fl uorescent ABPs MDW941 and MDW933. A: Detection of GBA in homogenates of cultured fi broblasts (30 g): labeling followed by SDS-PAGE and detection by fl uorescence imaging. AMRF 1, patient 1 (LIMP2 W178X/W178X); AMRF 2, patient 2 (LIMP2 W178X/W178X); AMRF 3, patient 3 (LIMP2 W178X/W178X); Car, carrier of AMRF (LIMP2 W178X/WT); GD, type 2 GD patient; Rec. GBA, recombinant GBA (Cerezyme). B: Fluorescence microscopy. Fibroblasts labeled in vivo with MDW933 and DAPI [Upper micrograph, control fi broblasts (WT); lower micrograph, AMRF patient]. The scale bar represents 20 m. C: Detection of GBA in homogenates of leukocytes (50 g): labeling followed by SDS-PAGE and detection by fl uorescence imaging. D: Detection of GBA in homog- enates of cultured macrophages (20 g): labeling followed by SDS-PAGE and detection by fl uorescence imaging.

Article Snippet: The rabbit polyclonal anti-LIMP2 antibody was purchased to Novus Biologicals, Littleton, CO.

Techniques: Cell Culture, Labeling, SDS Page, Imaging, Recombinant, Fluorescence, Microscopy, In Vivo, Control

Journal: Journal of Lipid Research

Article Title: Action myoclonus-renal failure syndrome: diagnostic applications of activity-based probes and lipid analysis

doi: 10.1194/jlr.m043802

Figure Lengend Snippet: Fig. 3. Partial secretion of GBA by fi broblasts. Fibroblasts were labeled for 72 h with MDW941. From 1 ml medium, GBA was im- munoprecipitated with anti-GBA monoclonal antibody (8E4) im- mobilized to Sepharose beads. The immunoprecipitate, subjected to SDS-PAGE and fl uorescent GBA labeled by MDW941, was visual- ized by imaging. Lane 1, carrier (Car) of AMRF (LIMP2 W178X/ WT); lane 2, AMRF patient 1 (AMRF 1) (LIMP2 W178X/W178X); lane 3, carrier of AMRF (LIMP2 W178X/WT); lane 4, AMRF pa- tient 3 (AMRF 3) (LIMP2 W178X/W178X); lane 5, control subject (WT) (LIMP2 WT); lane 6, recombinant GBA (Rec. GBA ).

Article Snippet: The rabbit polyclonal anti-LIMP2 antibody was purchased to Novus Biologicals, Littleton, CO.

Techniques: Labeling, SDS Page, Imaging, Control, Recombinant

Journal: Journal of Lipid Research

Article Title: Action myoclonus-renal failure syndrome: diagnostic applications of activity-based probes and lipid analysis

doi: 10.1194/jlr.m043802

Figure Lengend Snippet: Fig. 4. Glucosylceramide and glucosylsphingosine content of cells and plasma in relation to AMRF/LIMP2 defi ciency. Glucosylceramide and glucosylsphingosine were determined as described in Materials and Meth- ods. A: Glucosylceramide (nmol/mg total protein) in AMRF fi broblasts (n = 2). B: Glucosylceramide (nmol/ ml total protein) in human AMRF plasma specimens (n = 2). C: Glucosylsphingosine (pmol/mg total pro- tein) in AMRF fi broblasts (n = 3). D: Glucosylsphingosine (pmol/mg total protein) in AMRF leukocytes (n = 1). E: Glucosylsphingosine (pmol/ml) in human AMRF plasma specimens (n = 3). F: Glucosylsphingosine (pmol/ml) in samples from LIMP2 / mice (n = 3). AMRF, LIMP2-defi cient patients (LIMP2 W178X/ W178X); GD, GD patients; GD Car, GD carriers; LIMP2 +/+ , WT mice; LIMP2 / , KO LIMP2 mice. ** P < 0.01, unpaired student’s t -test ).

Article Snippet: The rabbit polyclonal anti-LIMP2 antibody was purchased to Novus Biologicals, Littleton, CO.

Techniques: Clinical Proteomics