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: Nature communications

Article Title: SCARB2 drives hepatocellular carcinoma tumor initiating cells via enhanced MYC transcriptional activity.

doi: 10.1038/s41467-023-41593-z

Figure Lengend Snippet: Fig. 6 | PMB binds with SCARB2 to suppress HCC by decreasing HDAC3- mediated MYC acetylation and MYC transcriptional activity. a–b Virtual screening of FDA-approved drugs to identify small molecules binding with SCARB2 (a), top 10 hits are listed (b). c The kinetics of the SCARB2-PMB interaction were determined by surface plasmon resonance (SPR) analysis. d The highest scoring docking model of the SCARB2 and PMB complex is shown. Top: surface of the PMB- SCARB2 complex. Bottom: 3D structure of the PMB (yellow)-SCARB2 complex. e The kinetics of the SCARB2-MYC interaction with or without PMB were deter- mined by SPR. f Structured illumination microscopic (SIM) images of vehicle- or PMB- treated HCCLM3 cells (1 h) stained for MYC and SCARB2. Scale bar, 10 μm. g The effect of PMB on the interaction of MYC and SCARB2 was evaluated by Co-IP assays. Extracts of DMSO and PMB-treated HCCLM3 cells were IP with an anti- SCARB2 Ab. h Representative images of MYC/HDAC3 colocalization foci in HCCLM3 cells before and after PMB treatment. Scale bar, 10 μm. i Effect of PMB on MYC acetylation. Extracts of DMSO and PMB-treated HCCLM3 cells were IP with an anti-MYC Ab. Acetylated MYC was detected by immunoblotting. j HCCLM3 cells

Article Snippet: Scarb2 knockout (Scarb2loxP/loxP, Scarb2F/F) (5–6 weeks old, 1 male and 2 females)micewere generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Activity Assay, Binding Assay, SPR Assay, Staining, Co-Immunoprecipitation Assay, Western Blot

Journal: Veterinary research

Article Title: Incomplete autophagy promotes the proliferation of Mycoplasma hyopneumoniae through the JNK and Akt pathways in porcine alveolar macrophages.

doi: 10.1186/s13567-022-01074-5

Figure Lengend Snippet: Figure 5 Colocalization of autophagosomes and lysosomes analysed by immunofluorescence. A Immunofluorescence of 3D4/21 cells that were infected with Ad-mRFP-GFP-LC3 and mock-infected, cultured in EBSS, treated with HCQ, or infected with AH at an MOI of 200 after 6 h. Thirty-two cells were randomly selected from each group; the yellow and red dots were counted for each cell. B Immunofluorescence of uninfected and infected PAMs collected from pigs and cultured in 24-well plates for 9 h. Yellow fluorescent puncta indicate the overlap of Mhp366 (green) and LAMP2 (red). Twenty cells were randomly selected from each group, and the yellow fluorescent puncta were counted from each cell. C Immunofluorescence of 3D4/21 cells cultured in EBSS medium or infected with AH after 6 h. Forty cells were randomly selected from each group, and the yellow fluorescent puncta were counted for each cell. Between-group differences were assessed using a t test; **p ≤ 0.01.

Article Snippet: Primary antibodies against β-actin (66009-1-Ig), p38 MAPK (66234-4-Ig), lysosome-associated membrane protein 2 (LAMP2) (66301-1), and extracellular signal-regulated kinases 1 and 2 (Erk1/2) (16443-1-AP) and fluorescent secondary antibodies (CoraLite594-conjugated donkey anti-rabbit IgG [SA00006-8] and Alexa Fluor 488-conjugated AffiniPure goat anti-mouse IgG [SA00006-1]) were obtained from Proteintech (Wuhan, China).

Techniques: Immunofluorescence, Infection, Cell Culture

Journal: Human Molecular Genetics

Article Title: Glucocerebrosidase modulates cognitive and motor activities in murine models of Parkinson’s disease

doi: 10.1093/hmg/ddw124

Figure Lengend Snippet: Effects of partial glucocerebrosidase inhibition on gene expression in the striata of WT and PrP-A53T-SNCA mice

Article Snippet: , LIMP-2 , Scarb2 , 1.00 ± 0.15 , 1.15 ± 0.26 , 1.05 ± 0.37 , 0.87 ± 0.08 , 12492 , Mm00446978_m1.

Techniques: Inhibition, Gene Expression, Saline, Control